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rabbit polyclonal anti ezrin antibody  (Developmental Studies Hybridoma Bank)
94
Developmental Studies Hybridoma Bank rabbit polyclonal anti ezrin antibody
A. Representative western blot from total cell lysates of MM6 and NOMO-1 cell lines demonstrating NOMO-1 cells have similar eIF4E levels to CD34+ from healthy donors, while MM6 cells have similar levels to high-eIF4E AML patient specimens. b-Actin is provided for loading control. Each lane refers to a different sample. B . Confocal micrograph of eIF4E nuclear and cytoplasmic staining in MM6 cells. Scale bar = 10 µm. C, D. Quantitation of western blots for NOMO-1 eIF4E relative to vector or MM6 CRISPR 4E relative to CRISPR-CTRL cells ( , Supplemental Figure 1G). Each data point represents a biological replicate. Bar represents the mean. Standard deviation and p-values (Welch’s t test) are shown. E. Representative confocal micrograph in NOMO-1 eIF4E cells demonstrating that HA staining is specific as its signal is removed upon treatment with hyaluronidase (HAse). HA is red; DAPI is blue. F. Representative western blot of MM6 cells treated with the eIF4E inhibitor ribavirin or vehicle control demonstrating that ribavirin reduces eIF4E target proteins including <t>Ezrin.</t> b-Actin is provided as a loading control. G, H . Representative western blots demonstrating lower eIF4E levels and factors in the Ezrin-CD44-HA axis in MM6 and THP-1 CRISPR-4E cells compared to CRISPR-Controls.
Rabbit Polyclonal Anti Ezrin Antibody, supplied by Developmental Studies Hybridoma Bank, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rabbit+polyclonal+anti+ezrin/anti-Ezrin/bio_rxiv__64898__2026__02__21__707190-344-200-213
Average 94 stars, based on 1 article reviews
rabbit polyclonal anti ezrin antibody - by Bioz Stars, 2026-09
94/100 stars
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rabbit polyclonal anti ezrin antibody  (Santa Cruz Biotechnology)
94
Santa Cruz Biotechnology rabbit polyclonal anti ezrin antibody
A. Representative western blot from total cell lysates of MM6 and NOMO-1 cell lines demonstrating NOMO-1 cells have similar eIF4E levels to CD34+ from healthy donors, while MM6 cells have similar levels to high-eIF4E AML patient specimens. b-Actin is provided for loading control. Each lane refers to a different sample. B . Confocal micrograph of eIF4E nuclear and cytoplasmic staining in MM6 cells. Scale bar = 10 µm. C, D. Quantitation of western blots for NOMO-1 eIF4E relative to vector or MM6 CRISPR 4E relative to CRISPR-CTRL cells ( , Supplemental Figure 1G). Each data point represents a biological replicate. Bar represents the mean. Standard deviation and p-values (Welch’s t test) are shown. E. Representative confocal micrograph in NOMO-1 eIF4E cells demonstrating that HA staining is specific as its signal is removed upon treatment with hyaluronidase (HAse). HA is red; DAPI is blue. F. Representative western blot of MM6 cells treated with the eIF4E inhibitor ribavirin or vehicle control demonstrating that ribavirin reduces eIF4E target proteins including <t>Ezrin.</t> b-Actin is provided as a loading control. G, H . Representative western blots demonstrating lower eIF4E levels and factors in the Ezrin-CD44-HA axis in MM6 and THP-1 CRISPR-4E cells compared to CRISPR-Controls.
Rabbit Polyclonal Anti Ezrin Antibody, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rabbit+polyclonal+anti+ezrin/Ezrin+Antibody/bio_rxiv__64898__2026__02__21__707190-344-200-219
Average 94 stars, based on 1 article reviews
rabbit polyclonal anti ezrin antibody - by Bioz Stars, 2026-09
94/100 stars
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uk ezrin rabbit polyclonal  (Cell Signaling Technology Inc)
96
Cell Signaling Technology Inc uk ezrin rabbit polyclonal
A. Representative western blot from total cell lysates of MM6 and NOMO-1 cell lines demonstrating NOMO-1 cells have similar eIF4E levels to CD34+ from healthy donors, while MM6 cells have similar levels to high-eIF4E AML patient specimens. b-Actin is provided for loading control. Each lane refers to a different sample. B . Confocal micrograph of eIF4E nuclear and cytoplasmic staining in MM6 cells. Scale bar = 10 µm. C, D. Quantitation of western blots for NOMO-1 eIF4E relative to vector or MM6 CRISPR 4E relative to CRISPR-CTRL cells ( , Supplemental Figure 1G). Each data point represents a biological replicate. Bar represents the mean. Standard deviation and p-values (Welch’s t test) are shown. E. Representative confocal micrograph in NOMO-1 eIF4E cells demonstrating that HA staining is specific as its signal is removed upon treatment with hyaluronidase (HAse). HA is red; DAPI is blue. F. Representative western blot of MM6 cells treated with the eIF4E inhibitor ribavirin or vehicle control demonstrating that ribavirin reduces eIF4E target proteins including <t>Ezrin.</t> b-Actin is provided as a loading control. G, H . Representative western blots demonstrating lower eIF4E levels and factors in the Ezrin-CD44-HA axis in MM6 and THP-1 CRISPR-4E cells compared to CRISPR-Controls.
Uk Ezrin Rabbit Polyclonal, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rabbit+polyclonal+anti+ezrin/Ezrin+Antibody/pmc12432371__mmc1-92-12-18
Average 96 stars, based on 1 article reviews
uk ezrin rabbit polyclonal - by Bioz Stars, 2026-09
96/100 stars
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polyclonal rabbit  (Cell Signaling Technology Inc)
96
Cell Signaling Technology Inc polyclonal rabbit
A. Representative western blot from total cell lysates of MM6 and NOMO-1 cell lines demonstrating NOMO-1 cells have similar eIF4E levels to CD34+ from healthy donors, while MM6 cells have similar levels to high-eIF4E AML patient specimens. b-Actin is provided for loading control. Each lane refers to a different sample. B . Confocal micrograph of eIF4E nuclear and cytoplasmic staining in MM6 cells. Scale bar = 10 µm. C, D. Quantitation of western blots for NOMO-1 eIF4E relative to vector or MM6 CRISPR 4E relative to CRISPR-CTRL cells ( , Supplemental Figure 1G). Each data point represents a biological replicate. Bar represents the mean. Standard deviation and p-values (Welch’s t test) are shown. E. Representative confocal micrograph in NOMO-1 eIF4E cells demonstrating that HA staining is specific as its signal is removed upon treatment with hyaluronidase (HAse). HA is red; DAPI is blue. F. Representative western blot of MM6 cells treated with the eIF4E inhibitor ribavirin or vehicle control demonstrating that ribavirin reduces eIF4E target proteins including <t>Ezrin.</t> b-Actin is provided as a loading control. G, H . Representative western blots demonstrating lower eIF4E levels and factors in the Ezrin-CD44-HA axis in MM6 and THP-1 CRISPR-4E cells compared to CRISPR-Controls.
Polyclonal Rabbit, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rabbit+polyclonal+anti+ezrin/Ezrin+Antibody/us12303617-564-12-19
Average 96 stars, based on 1 article reviews
polyclonal rabbit - by Bioz Stars, 2026-09
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rabbit anti phospho  (OriGene)
92
OriGene rabbit anti phospho
A. Representative western blot from total cell lysates of MM6 and NOMO-1 cell lines demonstrating NOMO-1 cells have similar eIF4E levels to CD34+ from healthy donors, while MM6 cells have similar levels to high-eIF4E AML patient specimens. b-Actin is provided for loading control. Each lane refers to a different sample. B . Confocal micrograph of eIF4E nuclear and cytoplasmic staining in MM6 cells. Scale bar = 10 µm. C, D. Quantitation of western blots for NOMO-1 eIF4E relative to vector or MM6 CRISPR 4E relative to CRISPR-CTRL cells ( , Supplemental Figure 1G). Each data point represents a biological replicate. Bar represents the mean. Standard deviation and p-values (Welch’s t test) are shown. E. Representative confocal micrograph in NOMO-1 eIF4E cells demonstrating that HA staining is specific as its signal is removed upon treatment with hyaluronidase (HAse). HA is red; DAPI is blue. F. Representative western blot of MM6 cells treated with the eIF4E inhibitor ribavirin or vehicle control demonstrating that ribavirin reduces eIF4E target proteins including <t>Ezrin.</t> b-Actin is provided as a loading control. G, H . Representative western blots demonstrating lower eIF4E levels and factors in the Ezrin-CD44-HA axis in MM6 and THP-1 CRISPR-4E cells compared to CRISPR-Controls.
Rabbit Anti Phospho, supplied by OriGene, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rabbit+polyclonal+anti+ezrin/Ezrin+(EZR)+Rabbit+Polyclonal+Antibody/pm39158743-119-12-17
Average 92 stars, based on 1 article reviews
rabbit anti phospho - by Bioz Stars, 2026-09
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mouse anti phospho  (OriGene)
92
OriGene mouse anti phospho
A. Representative western blot from total cell lysates of MM6 and NOMO-1 cell lines demonstrating NOMO-1 cells have similar eIF4E levels to CD34+ from healthy donors, while MM6 cells have similar levels to high-eIF4E AML patient specimens. b-Actin is provided for loading control. Each lane refers to a different sample. B . Confocal micrograph of eIF4E nuclear and cytoplasmic staining in MM6 cells. Scale bar = 10 µm. C, D. Quantitation of western blots for NOMO-1 eIF4E relative to vector or MM6 CRISPR 4E relative to CRISPR-CTRL cells ( , Supplemental Figure 1G). Each data point represents a biological replicate. Bar represents the mean. Standard deviation and p-values (Welch’s t test) are shown. E. Representative confocal micrograph in NOMO-1 eIF4E cells demonstrating that HA staining is specific as its signal is removed upon treatment with hyaluronidase (HAse). HA is red; DAPI is blue. F. Representative western blot of MM6 cells treated with the eIF4E inhibitor ribavirin or vehicle control demonstrating that ribavirin reduces eIF4E target proteins including <t>Ezrin.</t> b-Actin is provided as a loading control. G, H . Representative western blots demonstrating lower eIF4E levels and factors in the Ezrin-CD44-HA axis in MM6 and THP-1 CRISPR-4E cells compared to CRISPR-Controls.
Mouse Anti Phospho, supplied by OriGene, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rabbit+polyclonal+anti+ezrin/Ezrin+(EZR)+Rabbit+Polyclonal+Antibody/pmc11335284-88-25-33
Average 92 stars, based on 1 article reviews
mouse anti phospho - by Bioz Stars, 2026-09
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ezrin erm polyclonal rabbit antibody  (Cell Signaling Technology Inc)
95
Cell Signaling Technology Inc ezrin erm polyclonal rabbit antibody
A. Representative western blot from total cell lysates of MM6 and NOMO-1 cell lines demonstrating NOMO-1 cells have similar eIF4E levels to CD34+ from healthy donors, while MM6 cells have similar levels to high-eIF4E AML patient specimens. b-Actin is provided for loading control. Each lane refers to a different sample. B . Confocal micrograph of eIF4E nuclear and cytoplasmic staining in MM6 cells. Scale bar = 10 µm. C, D. Quantitation of western blots for NOMO-1 eIF4E relative to vector or MM6 CRISPR 4E relative to CRISPR-CTRL cells ( , Supplemental Figure 1G). Each data point represents a biological replicate. Bar represents the mean. Standard deviation and p-values (Welch’s t test) are shown. E. Representative confocal micrograph in NOMO-1 eIF4E cells demonstrating that HA staining is specific as its signal is removed upon treatment with hyaluronidase (HAse). HA is red; DAPI is blue. F. Representative western blot of MM6 cells treated with the eIF4E inhibitor ribavirin or vehicle control demonstrating that ribavirin reduces eIF4E target proteins including <t>Ezrin.</t> b-Actin is provided as a loading control. G, H . Representative western blots demonstrating lower eIF4E levels and factors in the Ezrin-CD44-HA axis in MM6 and THP-1 CRISPR-4E cells compared to CRISPR-Controls.
Ezrin Erm Polyclonal Rabbit Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rabbit+polyclonal+anti+ezrin/Ezrin%2FRadixin%2FMoesin+Antibody/pm38904854-115-8-14
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ezrin erm polyclonal rabbit antibody - by Bioz Stars, 2026-09
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rabbit polyclonal anti phospho erm  (Cell Signaling Technology Inc)
95
Cell Signaling Technology Inc rabbit polyclonal anti phospho erm

Rabbit Polyclonal Anti Phospho Erm, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rabbit+polyclonal+anti+ezrin/Ezrin%2FRadixin%2FMoesin+Antibody/pmc10959657-8-0-10
Average 95 stars, based on 1 article reviews
rabbit polyclonal anti phospho erm - by Bioz Stars, 2026-09
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Image Search Results


A. Representative western blot from total cell lysates of MM6 and NOMO-1 cell lines demonstrating NOMO-1 cells have similar eIF4E levels to CD34+ from healthy donors, while MM6 cells have similar levels to high-eIF4E AML patient specimens. b-Actin is provided for loading control. Each lane refers to a different sample. B . Confocal micrograph of eIF4E nuclear and cytoplasmic staining in MM6 cells. Scale bar = 10 µm. C, D. Quantitation of western blots for NOMO-1 eIF4E relative to vector or MM6 CRISPR 4E relative to CRISPR-CTRL cells ( , Supplemental Figure 1G). Each data point represents a biological replicate. Bar represents the mean. Standard deviation and p-values (Welch’s t test) are shown. E. Representative confocal micrograph in NOMO-1 eIF4E cells demonstrating that HA staining is specific as its signal is removed upon treatment with hyaluronidase (HAse). HA is red; DAPI is blue. F. Representative western blot of MM6 cells treated with the eIF4E inhibitor ribavirin or vehicle control demonstrating that ribavirin reduces eIF4E target proteins including Ezrin. b-Actin is provided as a loading control. G, H . Representative western blots demonstrating lower eIF4E levels and factors in the Ezrin-CD44-HA axis in MM6 and THP-1 CRISPR-4E cells compared to CRISPR-Controls.

Journal: bioRxiv

Article Title: eIF4E and Ezrin cooperate in pseudopods to drive a localized migratory translation program in acute myeloid leukemia

doi: 10.64898/2026.02.21.707190

Figure Lengend Snippet: A. Representative western blot from total cell lysates of MM6 and NOMO-1 cell lines demonstrating NOMO-1 cells have similar eIF4E levels to CD34+ from healthy donors, while MM6 cells have similar levels to high-eIF4E AML patient specimens. b-Actin is provided for loading control. Each lane refers to a different sample. B . Confocal micrograph of eIF4E nuclear and cytoplasmic staining in MM6 cells. Scale bar = 10 µm. C, D. Quantitation of western blots for NOMO-1 eIF4E relative to vector or MM6 CRISPR 4E relative to CRISPR-CTRL cells ( , Supplemental Figure 1G). Each data point represents a biological replicate. Bar represents the mean. Standard deviation and p-values (Welch’s t test) are shown. E. Representative confocal micrograph in NOMO-1 eIF4E cells demonstrating that HA staining is specific as its signal is removed upon treatment with hyaluronidase (HAse). HA is red; DAPI is blue. F. Representative western blot of MM6 cells treated with the eIF4E inhibitor ribavirin or vehicle control demonstrating that ribavirin reduces eIF4E target proteins including Ezrin. b-Actin is provided as a loading control. G, H . Representative western blots demonstrating lower eIF4E levels and factors in the Ezrin-CD44-HA axis in MM6 and THP-1 CRISPR-4E cells compared to CRISPR-Controls.

Article Snippet: Antibodies for immunoblotting: mouse monoclonal anti-eIF4E (cat# 610270, BD Biosciences), mouse monoclonal anti-β-Actin (cat# A5441, Sigma Aldrich), rabbit polyclonal anti-Mcl-1 (S-19) (cat# sc-819, Santa Cruz), mouse monoclonal anti-HSP90α/β (F-8) (cat# sc-13119, Santa Cruz), rabbit polyclonal anti-Myc (cat# ab32072, Abcam), Mouse monoclonal anti-CD44 antibody (cat# 156–3 C11, Cell Signaling Technology), rabbit polyclonal anti-CD44 (cat# A12410, Abclonal), rabbit polyclonal anti-HAS3 antibody (cat# ab154104, Abcam), rabbit polyclonal anti-phosphoglucomutase 5 (cat# AI14638, Abgent), rabbit polyclonal anti-Lamin A (C-terminal) (cat# L1293, Sigma Aldrich), rabbit polyclonal anti-GAPDH (FL-335) (cat# sc-25778, Santa Cruz), rabbit polyclonal anti-UGDH (cat#AP12613b-EV, Abgent), rabbit monoclonal anti-Hexokinase II (cat# 2867, Cell Signaling Technology), rabbit monoclonal anti-UAP1 antibody (cat# 2716, GenuinBiotech), mouse monoclonal anti-RPL17 (C-8, cat# sc-515904, Santa Cruz), mouse monoclonal anti-RPS16 (D-8, cat# sc-518206; Santa Cruz), mouse monoclonal anti-RPS6 (C-8, cat# sc-74459, Santa Cruz), mouse monoclonal anti-eIF3e (G-7, cat# sc-390413, Santa Cruz), mouse monoclonal anti-eIF3p110 (B-6, cat# sc-74507, Santa Cruz), mouse monoclonal anti-eIF3b (A-7, cat# sc-374156, Santa Cruz), mouse monoclonal anti-eIF4AI/II (H-5, cat# sc-377315, Santa Cruz), mouse monoclonal anti-eIF4G (A-10, cat# sc-133155, Santa Cruz), mouse monoclonal anti-Nopp140 (E-7, cat# sc-374033, Santa Cruz), rabbit monoclonal anti-histone H3 acetyl K27 (cat# ab177178, Abcam), rabbit polyclonal Calreticulin (cat# AW5211, ABCEPTA), rabbit polyclonal anti-Calnexin (cat# ab22595, Abcam), rabbit polyclonal anti-Ezrin antibody (cat# PA5-80603, Invitrogen), mouse monoclonal anti-Ezrin (CPTC-Ezrin-1, cat# AB_2100318, DSHB), mouse monoclonal anti-MEK-1 (H-8, sc-6250, Santa Cruz), and mouse monoclonal anti-cytochrome c (A-8, cat# sc-13156, Santa Cruz).

Techniques: Western Blot, Control, Staining, Quantitation Assay, Plasmid Preparation, CRISPR, Standard Deviation

B. The impact on the Ezrin-CD44-HA axis of NOMO-1 eIF4E overexpression (eIF4E) compared to vector control (A) or of MM6 CRISPR-4E cells relative to MM6 CRISPR-CTRL cells (B). Left panels. Western blots of eIF4E and Ezrin-CD44-HA axis. Corresponding b-Actin shown for loading controls. Right panels . Confocal micrographs (single section through the plane of cells) of the indicated cells stained for HA or CD44 shown in red. DAPI shown in blue. Scale bar = 10µm. C. Western blot of primary specimens from AML patients (with normal or high-eIF4E) and CD34 + from healthy donors showing expression of eIF4E and Ezrin. b-Actin is provided as a loading control. Each lane represents a different individual. D. Adhesion (left) and invasion (right) capacity using HS-5 stromal cells of NOMO-1 and MM6 cell lines as a function of genetic eIF4E overexpression or CRISPR knockdown respectively. Data is presented as fold change relative to their corresponding controls. Each symbol represents an independent experiment performed with replicates. The bar represents the mean with standard deviations and p-values calculated with two tailed paired T test. E. Schematic representation of colonization assay into mesenchymal stromal cell spheroid model mimicking the bone marrow niche. F. Quantification of AML cell colonization capacity relative to their corresponding controls. Each symbol represents an experimental replicate. The bar represents the mean with standard deviations and p-values, Welch’s t test.

Journal: bioRxiv

Article Title: eIF4E and Ezrin cooperate in pseudopods to drive a localized migratory translation program in acute myeloid leukemia

doi: 10.64898/2026.02.21.707190

Figure Lengend Snippet: B. The impact on the Ezrin-CD44-HA axis of NOMO-1 eIF4E overexpression (eIF4E) compared to vector control (A) or of MM6 CRISPR-4E cells relative to MM6 CRISPR-CTRL cells (B). Left panels. Western blots of eIF4E and Ezrin-CD44-HA axis. Corresponding b-Actin shown for loading controls. Right panels . Confocal micrographs (single section through the plane of cells) of the indicated cells stained for HA or CD44 shown in red. DAPI shown in blue. Scale bar = 10µm. C. Western blot of primary specimens from AML patients (with normal or high-eIF4E) and CD34 + from healthy donors showing expression of eIF4E and Ezrin. b-Actin is provided as a loading control. Each lane represents a different individual. D. Adhesion (left) and invasion (right) capacity using HS-5 stromal cells of NOMO-1 and MM6 cell lines as a function of genetic eIF4E overexpression or CRISPR knockdown respectively. Data is presented as fold change relative to their corresponding controls. Each symbol represents an independent experiment performed with replicates. The bar represents the mean with standard deviations and p-values calculated with two tailed paired T test. E. Schematic representation of colonization assay into mesenchymal stromal cell spheroid model mimicking the bone marrow niche. F. Quantification of AML cell colonization capacity relative to their corresponding controls. Each symbol represents an experimental replicate. The bar represents the mean with standard deviations and p-values, Welch’s t test.

Article Snippet: Antibodies for immunoblotting: mouse monoclonal anti-eIF4E (cat# 610270, BD Biosciences), mouse monoclonal anti-β-Actin (cat# A5441, Sigma Aldrich), rabbit polyclonal anti-Mcl-1 (S-19) (cat# sc-819, Santa Cruz), mouse monoclonal anti-HSP90α/β (F-8) (cat# sc-13119, Santa Cruz), rabbit polyclonal anti-Myc (cat# ab32072, Abcam), Mouse monoclonal anti-CD44 antibody (cat# 156–3 C11, Cell Signaling Technology), rabbit polyclonal anti-CD44 (cat# A12410, Abclonal), rabbit polyclonal anti-HAS3 antibody (cat# ab154104, Abcam), rabbit polyclonal anti-phosphoglucomutase 5 (cat# AI14638, Abgent), rabbit polyclonal anti-Lamin A (C-terminal) (cat# L1293, Sigma Aldrich), rabbit polyclonal anti-GAPDH (FL-335) (cat# sc-25778, Santa Cruz), rabbit polyclonal anti-UGDH (cat#AP12613b-EV, Abgent), rabbit monoclonal anti-Hexokinase II (cat# 2867, Cell Signaling Technology), rabbit monoclonal anti-UAP1 antibody (cat# 2716, GenuinBiotech), mouse monoclonal anti-RPL17 (C-8, cat# sc-515904, Santa Cruz), mouse monoclonal anti-RPS16 (D-8, cat# sc-518206; Santa Cruz), mouse monoclonal anti-RPS6 (C-8, cat# sc-74459, Santa Cruz), mouse monoclonal anti-eIF3e (G-7, cat# sc-390413, Santa Cruz), mouse monoclonal anti-eIF3p110 (B-6, cat# sc-74507, Santa Cruz), mouse monoclonal anti-eIF3b (A-7, cat# sc-374156, Santa Cruz), mouse monoclonal anti-eIF4AI/II (H-5, cat# sc-377315, Santa Cruz), mouse monoclonal anti-eIF4G (A-10, cat# sc-133155, Santa Cruz), mouse monoclonal anti-Nopp140 (E-7, cat# sc-374033, Santa Cruz), rabbit monoclonal anti-histone H3 acetyl K27 (cat# ab177178, Abcam), rabbit polyclonal Calreticulin (cat# AW5211, ABCEPTA), rabbit polyclonal anti-Calnexin (cat# ab22595, Abcam), rabbit polyclonal anti-Ezrin antibody (cat# PA5-80603, Invitrogen), mouse monoclonal anti-Ezrin (CPTC-Ezrin-1, cat# AB_2100318, DSHB), mouse monoclonal anti-MEK-1 (H-8, sc-6250, Santa Cruz), and mouse monoclonal anti-cytochrome c (A-8, cat# sc-13156, Santa Cruz).

Techniques: Over Expression, Plasmid Preparation, Control, CRISPR, Western Blot, Staining, Expressing, Knockdown, Two Tailed Test

A. Western blot of total cell lysates from MM6 cells grown in suspension demonstrated knockdown of Ezrin (siEZR) or eIF4E (siEIF4E) compared to RNAi to luciferase (siLUC) used as a negative control. b-Actin is provided as a loading control. Other proteins of the Ezrin-CD44-HA axis are also shown. Quantification for these is shown in with 3-6 biological replicates for each protein. B. Adhesion and invasion capacity of MM6 cells onto/through HS-5 bone marrow stroma. Fold change relative to siLUC is shown. Each symbol represents a biological replicate performed independently with replicates. Bars represent the mean, shown with standard deviations and p-values (one-way ANOVA). C. Western blot of eIF4E and Ezrin immunoprecipitations (IPs) using total cell lysates from MM6 cells in suspension. SN, supernatant after immunoprecipitation, IgG, negative control. Representative of three biological replicates. IPs of total cell lysates from THP-1 cells in suspension are provided in . D. IPs from MM6 cells in suspension using the rRNA antibody Y10b. LC indicates antibody light chain. Representative of three biological replicates. E. RNA immunoprecipitations (RIPs) from MM6 total cell lysates grown in suspension using anti-Ezrin (Ezrin RIP) or anti-eIF4E (eIF4E RIP) antibodies. Data are from RT-qPCR and represented relative to input. Each symbol represents a biological replicate performed independently with triplicates. Bars represent the mean, shown with standard deviations and p-values (two tailed Welch’s t test). F. Western blots of eIF4E and Ezrin IPs from the cytoplasmic fractions of MM6 cells in suspension or G . After invasion through HS-5 bone marrow stroma (invaded). Fractionation controls are provided in . H. RIPs from MM6 cytoplasmic fraction from invaded cells using anti-eIF4E (eIF4E RIP) or anti-Ezrin (Ezrin RIP) antibodies. Data are from RT-qPCR represented relative to input. Each symbol represents a biological replicate performed independently with triplicates. Bars represent the mean, shown with standard deviations and p-values (two tailed Welch’s t test). I. Count of the number of pseudopods observed in suspension and invaded MM6 cells represented as a fraction relative to the total cells counted. Each symbol represents a biological replicate. Bars represent the mean, shown with standard deviations and p-values (two-way ANOVA). J . Immunofluorescence and confocal microscopy demonstrating eIF4E, Ezrin, CD44 and rRNA are localized to the same pseudopods (white arrows). All confocal micrographs represent a single section through the plane of the cell. Scale bar = 10 µm

Journal: bioRxiv

Article Title: eIF4E and Ezrin cooperate in pseudopods to drive a localized migratory translation program in acute myeloid leukemia

doi: 10.64898/2026.02.21.707190

Figure Lengend Snippet: A. Western blot of total cell lysates from MM6 cells grown in suspension demonstrated knockdown of Ezrin (siEZR) or eIF4E (siEIF4E) compared to RNAi to luciferase (siLUC) used as a negative control. b-Actin is provided as a loading control. Other proteins of the Ezrin-CD44-HA axis are also shown. Quantification for these is shown in with 3-6 biological replicates for each protein. B. Adhesion and invasion capacity of MM6 cells onto/through HS-5 bone marrow stroma. Fold change relative to siLUC is shown. Each symbol represents a biological replicate performed independently with replicates. Bars represent the mean, shown with standard deviations and p-values (one-way ANOVA). C. Western blot of eIF4E and Ezrin immunoprecipitations (IPs) using total cell lysates from MM6 cells in suspension. SN, supernatant after immunoprecipitation, IgG, negative control. Representative of three biological replicates. IPs of total cell lysates from THP-1 cells in suspension are provided in . D. IPs from MM6 cells in suspension using the rRNA antibody Y10b. LC indicates antibody light chain. Representative of three biological replicates. E. RNA immunoprecipitations (RIPs) from MM6 total cell lysates grown in suspension using anti-Ezrin (Ezrin RIP) or anti-eIF4E (eIF4E RIP) antibodies. Data are from RT-qPCR and represented relative to input. Each symbol represents a biological replicate performed independently with triplicates. Bars represent the mean, shown with standard deviations and p-values (two tailed Welch’s t test). F. Western blots of eIF4E and Ezrin IPs from the cytoplasmic fractions of MM6 cells in suspension or G . After invasion through HS-5 bone marrow stroma (invaded). Fractionation controls are provided in . H. RIPs from MM6 cytoplasmic fraction from invaded cells using anti-eIF4E (eIF4E RIP) or anti-Ezrin (Ezrin RIP) antibodies. Data are from RT-qPCR represented relative to input. Each symbol represents a biological replicate performed independently with triplicates. Bars represent the mean, shown with standard deviations and p-values (two tailed Welch’s t test). I. Count of the number of pseudopods observed in suspension and invaded MM6 cells represented as a fraction relative to the total cells counted. Each symbol represents a biological replicate. Bars represent the mean, shown with standard deviations and p-values (two-way ANOVA). J . Immunofluorescence and confocal microscopy demonstrating eIF4E, Ezrin, CD44 and rRNA are localized to the same pseudopods (white arrows). All confocal micrographs represent a single section through the plane of the cell. Scale bar = 10 µm

Article Snippet: Antibodies for immunoblotting: mouse monoclonal anti-eIF4E (cat# 610270, BD Biosciences), mouse monoclonal anti-β-Actin (cat# A5441, Sigma Aldrich), rabbit polyclonal anti-Mcl-1 (S-19) (cat# sc-819, Santa Cruz), mouse monoclonal anti-HSP90α/β (F-8) (cat# sc-13119, Santa Cruz), rabbit polyclonal anti-Myc (cat# ab32072, Abcam), Mouse monoclonal anti-CD44 antibody (cat# 156–3 C11, Cell Signaling Technology), rabbit polyclonal anti-CD44 (cat# A12410, Abclonal), rabbit polyclonal anti-HAS3 antibody (cat# ab154104, Abcam), rabbit polyclonal anti-phosphoglucomutase 5 (cat# AI14638, Abgent), rabbit polyclonal anti-Lamin A (C-terminal) (cat# L1293, Sigma Aldrich), rabbit polyclonal anti-GAPDH (FL-335) (cat# sc-25778, Santa Cruz), rabbit polyclonal anti-UGDH (cat#AP12613b-EV, Abgent), rabbit monoclonal anti-Hexokinase II (cat# 2867, Cell Signaling Technology), rabbit monoclonal anti-UAP1 antibody (cat# 2716, GenuinBiotech), mouse monoclonal anti-RPL17 (C-8, cat# sc-515904, Santa Cruz), mouse monoclonal anti-RPS16 (D-8, cat# sc-518206; Santa Cruz), mouse monoclonal anti-RPS6 (C-8, cat# sc-74459, Santa Cruz), mouse monoclonal anti-eIF3e (G-7, cat# sc-390413, Santa Cruz), mouse monoclonal anti-eIF3p110 (B-6, cat# sc-74507, Santa Cruz), mouse monoclonal anti-eIF3b (A-7, cat# sc-374156, Santa Cruz), mouse monoclonal anti-eIF4AI/II (H-5, cat# sc-377315, Santa Cruz), mouse monoclonal anti-eIF4G (A-10, cat# sc-133155, Santa Cruz), mouse monoclonal anti-Nopp140 (E-7, cat# sc-374033, Santa Cruz), rabbit monoclonal anti-histone H3 acetyl K27 (cat# ab177178, Abcam), rabbit polyclonal Calreticulin (cat# AW5211, ABCEPTA), rabbit polyclonal anti-Calnexin (cat# ab22595, Abcam), rabbit polyclonal anti-Ezrin antibody (cat# PA5-80603, Invitrogen), mouse monoclonal anti-Ezrin (CPTC-Ezrin-1, cat# AB_2100318, DSHB), mouse monoclonal anti-MEK-1 (H-8, sc-6250, Santa Cruz), and mouse monoclonal anti-cytochrome c (A-8, cat# sc-13156, Santa Cruz).

Techniques: Western Blot, Suspension, Knockdown, Luciferase, Negative Control, Control, Immunoprecipitation, Quantitative RT-PCR, Two Tailed Test, Fractionation, Immunofluorescence, Confocal Microscopy

A. Genetic reduction of Ezrin and eIF4E using siRNA in MM6 cells grown in suspension impacts production of factors in the Ezrin-CD44-HA axis compared to the siLUC control shown in a representative western blot (left panel). b-Actin is provided as loading control. Right panel, quantification of protein expression for the indicated siRNAs relative to siLUC is shown. The expression of each protein was calculated relative to b-Actin. Each symbol represents a biological replicate. Means, standard deviations and p-values (multiple paired t tests). B. Endogenous Ezrin and eIF4E immunoprecipitations (IP) in THP-1 total cell lysates show similar observations to MM6 cells . SN supernatant after immunoprecipitation, IgG negative control. H2B also serves as a negative control for eIF4E and Ezrin IPs. C. Cytoplasmic (left) and total (right) cell lysates demonstrate Ezrin immunoprecipitated with the translation machinery including eIF4AI/II. D. Example fractionation controls for suspension (left) or invaded (right) cells shown in indicating quality of fractions. MEK and CalR are cytoplasmic markers; NOPP140 and H3K27A are nuclear markers.

Journal: bioRxiv

Article Title: eIF4E and Ezrin cooperate in pseudopods to drive a localized migratory translation program in acute myeloid leukemia

doi: 10.64898/2026.02.21.707190

Figure Lengend Snippet: A. Genetic reduction of Ezrin and eIF4E using siRNA in MM6 cells grown in suspension impacts production of factors in the Ezrin-CD44-HA axis compared to the siLUC control shown in a representative western blot (left panel). b-Actin is provided as loading control. Right panel, quantification of protein expression for the indicated siRNAs relative to siLUC is shown. The expression of each protein was calculated relative to b-Actin. Each symbol represents a biological replicate. Means, standard deviations and p-values (multiple paired t tests). B. Endogenous Ezrin and eIF4E immunoprecipitations (IP) in THP-1 total cell lysates show similar observations to MM6 cells . SN supernatant after immunoprecipitation, IgG negative control. H2B also serves as a negative control for eIF4E and Ezrin IPs. C. Cytoplasmic (left) and total (right) cell lysates demonstrate Ezrin immunoprecipitated with the translation machinery including eIF4AI/II. D. Example fractionation controls for suspension (left) or invaded (right) cells shown in indicating quality of fractions. MEK and CalR are cytoplasmic markers; NOPP140 and H3K27A are nuclear markers.

Article Snippet: Antibodies for immunoblotting: mouse monoclonal anti-eIF4E (cat# 610270, BD Biosciences), mouse monoclonal anti-β-Actin (cat# A5441, Sigma Aldrich), rabbit polyclonal anti-Mcl-1 (S-19) (cat# sc-819, Santa Cruz), mouse monoclonal anti-HSP90α/β (F-8) (cat# sc-13119, Santa Cruz), rabbit polyclonal anti-Myc (cat# ab32072, Abcam), Mouse monoclonal anti-CD44 antibody (cat# 156–3 C11, Cell Signaling Technology), rabbit polyclonal anti-CD44 (cat# A12410, Abclonal), rabbit polyclonal anti-HAS3 antibody (cat# ab154104, Abcam), rabbit polyclonal anti-phosphoglucomutase 5 (cat# AI14638, Abgent), rabbit polyclonal anti-Lamin A (C-terminal) (cat# L1293, Sigma Aldrich), rabbit polyclonal anti-GAPDH (FL-335) (cat# sc-25778, Santa Cruz), rabbit polyclonal anti-UGDH (cat#AP12613b-EV, Abgent), rabbit monoclonal anti-Hexokinase II (cat# 2867, Cell Signaling Technology), rabbit monoclonal anti-UAP1 antibody (cat# 2716, GenuinBiotech), mouse monoclonal anti-RPL17 (C-8, cat# sc-515904, Santa Cruz), mouse monoclonal anti-RPS16 (D-8, cat# sc-518206; Santa Cruz), mouse monoclonal anti-RPS6 (C-8, cat# sc-74459, Santa Cruz), mouse monoclonal anti-eIF3e (G-7, cat# sc-390413, Santa Cruz), mouse monoclonal anti-eIF3p110 (B-6, cat# sc-74507, Santa Cruz), mouse monoclonal anti-eIF3b (A-7, cat# sc-374156, Santa Cruz), mouse monoclonal anti-eIF4AI/II (H-5, cat# sc-377315, Santa Cruz), mouse monoclonal anti-eIF4G (A-10, cat# sc-133155, Santa Cruz), mouse monoclonal anti-Nopp140 (E-7, cat# sc-374033, Santa Cruz), rabbit monoclonal anti-histone H3 acetyl K27 (cat# ab177178, Abcam), rabbit polyclonal Calreticulin (cat# AW5211, ABCEPTA), rabbit polyclonal anti-Calnexin (cat# ab22595, Abcam), rabbit polyclonal anti-Ezrin antibody (cat# PA5-80603, Invitrogen), mouse monoclonal anti-Ezrin (CPTC-Ezrin-1, cat# AB_2100318, DSHB), mouse monoclonal anti-MEK-1 (H-8, sc-6250, Santa Cruz), and mouse monoclonal anti-cytochrome c (A-8, cat# sc-13156, Santa Cruz).

Techniques: Suspension, Control, Western Blot, Expressing, Immunoprecipitation, Negative Control, Fractionation

A. Western blots of total cell lysates for MM6 cells in suspension treated with puromycin and analyzed by WB with anti-puromycin antibody (to assess global protein synthesis) reveal that Ezrin does not influence global translation: RNAi knockdown to Ezrin (siEZR) did not show difference compared to luciferase (siLUC) control (left panel). In contrast, CRISPR 4E reduces global translation signal by about 50% relative to CRISPR-CTRL, consistent with previous studies. Each symbol represents a biological replicate, bar showing mean, standard deviation, p-value (two tailed paired t test). B. Confocal micrographs for VISTA-R and the indicated proteins for MM6 cells upon genetic knockdowns siEIF4E), siEZR or siLUC control Arrows indicate T-PODs. Scale bar = 10µm. Western blots confirmed knockdown (Supplementary Figure 6C) C. As in B but monitoring eIF4E in T-PODs as a function of Ezrin and eIF4E knockdown. Ezrin is also reduced by eIF4E knockdown in invaded cells (Supplementary Figure 6C). Scale bar = 10µm. D. Quantitation of intensity of factors as noted in T-PODs over 3 biological replicates for VISTA-R and rRNA, 5 for Ezrin and 2 for eIF4E. Boxplots show Imaris-quantified TPOD-associated intensities of VISTA-R, rRNA, Ezrin, and eIF4E under the conditions shown. Each dot represents an individual T-POD object, p-values calculated using one way ANOVA test.

Journal: bioRxiv

Article Title: eIF4E and Ezrin cooperate in pseudopods to drive a localized migratory translation program in acute myeloid leukemia

doi: 10.64898/2026.02.21.707190

Figure Lengend Snippet: A. Western blots of total cell lysates for MM6 cells in suspension treated with puromycin and analyzed by WB with anti-puromycin antibody (to assess global protein synthesis) reveal that Ezrin does not influence global translation: RNAi knockdown to Ezrin (siEZR) did not show difference compared to luciferase (siLUC) control (left panel). In contrast, CRISPR 4E reduces global translation signal by about 50% relative to CRISPR-CTRL, consistent with previous studies. Each symbol represents a biological replicate, bar showing mean, standard deviation, p-value (two tailed paired t test). B. Confocal micrographs for VISTA-R and the indicated proteins for MM6 cells upon genetic knockdowns siEIF4E), siEZR or siLUC control Arrows indicate T-PODs. Scale bar = 10µm. Western blots confirmed knockdown (Supplementary Figure 6C) C. As in B but monitoring eIF4E in T-PODs as a function of Ezrin and eIF4E knockdown. Ezrin is also reduced by eIF4E knockdown in invaded cells (Supplementary Figure 6C). Scale bar = 10µm. D. Quantitation of intensity of factors as noted in T-PODs over 3 biological replicates for VISTA-R and rRNA, 5 for Ezrin and 2 for eIF4E. Boxplots show Imaris-quantified TPOD-associated intensities of VISTA-R, rRNA, Ezrin, and eIF4E under the conditions shown. Each dot represents an individual T-POD object, p-values calculated using one way ANOVA test.

Article Snippet: Antibodies for immunoblotting: mouse monoclonal anti-eIF4E (cat# 610270, BD Biosciences), mouse monoclonal anti-β-Actin (cat# A5441, Sigma Aldrich), rabbit polyclonal anti-Mcl-1 (S-19) (cat# sc-819, Santa Cruz), mouse monoclonal anti-HSP90α/β (F-8) (cat# sc-13119, Santa Cruz), rabbit polyclonal anti-Myc (cat# ab32072, Abcam), Mouse monoclonal anti-CD44 antibody (cat# 156–3 C11, Cell Signaling Technology), rabbit polyclonal anti-CD44 (cat# A12410, Abclonal), rabbit polyclonal anti-HAS3 antibody (cat# ab154104, Abcam), rabbit polyclonal anti-phosphoglucomutase 5 (cat# AI14638, Abgent), rabbit polyclonal anti-Lamin A (C-terminal) (cat# L1293, Sigma Aldrich), rabbit polyclonal anti-GAPDH (FL-335) (cat# sc-25778, Santa Cruz), rabbit polyclonal anti-UGDH (cat#AP12613b-EV, Abgent), rabbit monoclonal anti-Hexokinase II (cat# 2867, Cell Signaling Technology), rabbit monoclonal anti-UAP1 antibody (cat# 2716, GenuinBiotech), mouse monoclonal anti-RPL17 (C-8, cat# sc-515904, Santa Cruz), mouse monoclonal anti-RPS16 (D-8, cat# sc-518206; Santa Cruz), mouse monoclonal anti-RPS6 (C-8, cat# sc-74459, Santa Cruz), mouse monoclonal anti-eIF3e (G-7, cat# sc-390413, Santa Cruz), mouse monoclonal anti-eIF3p110 (B-6, cat# sc-74507, Santa Cruz), mouse monoclonal anti-eIF3b (A-7, cat# sc-374156, Santa Cruz), mouse monoclonal anti-eIF4AI/II (H-5, cat# sc-377315, Santa Cruz), mouse monoclonal anti-eIF4G (A-10, cat# sc-133155, Santa Cruz), mouse monoclonal anti-Nopp140 (E-7, cat# sc-374033, Santa Cruz), rabbit monoclonal anti-histone H3 acetyl K27 (cat# ab177178, Abcam), rabbit polyclonal Calreticulin (cat# AW5211, ABCEPTA), rabbit polyclonal anti-Calnexin (cat# ab22595, Abcam), rabbit polyclonal anti-Ezrin antibody (cat# PA5-80603, Invitrogen), mouse monoclonal anti-Ezrin (CPTC-Ezrin-1, cat# AB_2100318, DSHB), mouse monoclonal anti-MEK-1 (H-8, sc-6250, Santa Cruz), and mouse monoclonal anti-cytochrome c (A-8, cat# sc-13156, Santa Cruz).

Techniques: Western Blot, Suspension, Knockdown, Luciferase, Control, CRISPR, Standard Deviation, Two Tailed Test, Quantitation Assay

A . Schematic comparing previous strategies to monitor in situ translation based on surrogate markers of translation such as labelled amino acids (left), and our novel method for direct measurement of translation, VISTA-R (right). B. VISTA-R signals in MM6 cells are directly related to translation activity since these are abrogated upon addition of translation inhibitors. Puromycin and/or HHT were added prior to the fixation step in VISTA-R. Representative confocal micrograph of VISTA-R signal in vehicle control, puromycin, HHT or combination treated cells. DAPI is shown in blue. Scale bar = 10 µm. C. Quantitation showing means, standard deviations of the mean, data points from each measurement, p-values calculated with one way ANOVA test from over 60 cells. D. T-PODs (as shown by arrows) contain active ribosomes as seen by VISTA-R (green). VISTA-R and rRNA (red), Ezrin (blue) overlay in these T-POD. E. Same as D but demonstrating VISTA-R (green), Ezrin (magenta) and RPS6 (red) and CD44 (turquoise) are present in the same T-PODs (see arrows). F. Same as D-E but demonstrating that VISTA-R (green), eIF4E (red) and Ezrin (magenta) and CD44 (turquoise) are present in the same T-PODs (see arrows). All confocal micrographs represent a single section through the plane of the cell. Scale bars = 10 µm.

Journal: bioRxiv

Article Title: eIF4E and Ezrin cooperate in pseudopods to drive a localized migratory translation program in acute myeloid leukemia

doi: 10.64898/2026.02.21.707190

Figure Lengend Snippet: A . Schematic comparing previous strategies to monitor in situ translation based on surrogate markers of translation such as labelled amino acids (left), and our novel method for direct measurement of translation, VISTA-R (right). B. VISTA-R signals in MM6 cells are directly related to translation activity since these are abrogated upon addition of translation inhibitors. Puromycin and/or HHT were added prior to the fixation step in VISTA-R. Representative confocal micrograph of VISTA-R signal in vehicle control, puromycin, HHT or combination treated cells. DAPI is shown in blue. Scale bar = 10 µm. C. Quantitation showing means, standard deviations of the mean, data points from each measurement, p-values calculated with one way ANOVA test from over 60 cells. D. T-PODs (as shown by arrows) contain active ribosomes as seen by VISTA-R (green). VISTA-R and rRNA (red), Ezrin (blue) overlay in these T-POD. E. Same as D but demonstrating VISTA-R (green), Ezrin (magenta) and RPS6 (red) and CD44 (turquoise) are present in the same T-PODs (see arrows). F. Same as D-E but demonstrating that VISTA-R (green), eIF4E (red) and Ezrin (magenta) and CD44 (turquoise) are present in the same T-PODs (see arrows). All confocal micrographs represent a single section through the plane of the cell. Scale bars = 10 µm.

Article Snippet: Antibodies for immunoblotting: mouse monoclonal anti-eIF4E (cat# 610270, BD Biosciences), mouse monoclonal anti-β-Actin (cat# A5441, Sigma Aldrich), rabbit polyclonal anti-Mcl-1 (S-19) (cat# sc-819, Santa Cruz), mouse monoclonal anti-HSP90α/β (F-8) (cat# sc-13119, Santa Cruz), rabbit polyclonal anti-Myc (cat# ab32072, Abcam), Mouse monoclonal anti-CD44 antibody (cat# 156–3 C11, Cell Signaling Technology), rabbit polyclonal anti-CD44 (cat# A12410, Abclonal), rabbit polyclonal anti-HAS3 antibody (cat# ab154104, Abcam), rabbit polyclonal anti-phosphoglucomutase 5 (cat# AI14638, Abgent), rabbit polyclonal anti-Lamin A (C-terminal) (cat# L1293, Sigma Aldrich), rabbit polyclonal anti-GAPDH (FL-335) (cat# sc-25778, Santa Cruz), rabbit polyclonal anti-UGDH (cat#AP12613b-EV, Abgent), rabbit monoclonal anti-Hexokinase II (cat# 2867, Cell Signaling Technology), rabbit monoclonal anti-UAP1 antibody (cat# 2716, GenuinBiotech), mouse monoclonal anti-RPL17 (C-8, cat# sc-515904, Santa Cruz), mouse monoclonal anti-RPS16 (D-8, cat# sc-518206; Santa Cruz), mouse monoclonal anti-RPS6 (C-8, cat# sc-74459, Santa Cruz), mouse monoclonal anti-eIF3e (G-7, cat# sc-390413, Santa Cruz), mouse monoclonal anti-eIF3p110 (B-6, cat# sc-74507, Santa Cruz), mouse monoclonal anti-eIF3b (A-7, cat# sc-374156, Santa Cruz), mouse monoclonal anti-eIF4AI/II (H-5, cat# sc-377315, Santa Cruz), mouse monoclonal anti-eIF4G (A-10, cat# sc-133155, Santa Cruz), mouse monoclonal anti-Nopp140 (E-7, cat# sc-374033, Santa Cruz), rabbit monoclonal anti-histone H3 acetyl K27 (cat# ab177178, Abcam), rabbit polyclonal Calreticulin (cat# AW5211, ABCEPTA), rabbit polyclonal anti-Calnexin (cat# ab22595, Abcam), rabbit polyclonal anti-Ezrin antibody (cat# PA5-80603, Invitrogen), mouse monoclonal anti-Ezrin (CPTC-Ezrin-1, cat# AB_2100318, DSHB), mouse monoclonal anti-MEK-1 (H-8, sc-6250, Santa Cruz), and mouse monoclonal anti-cytochrome c (A-8, cat# sc-13156, Santa Cruz).

Techniques: In Situ, Activity Assay, Control, Quantitation Assay

A. Schema of VISTA-R for invaded cells. Steps for isolated invaded AML cells for marking active ribosomes with the VISTA-R method. B. Confocal microscopy for patient specimen, AML23, demonstrated active translation, characterized by VISTA-R and Ezrin are present in the same pseudopods (white arrow). A single section through the plane of the cell is shown. Scale bar = 10µm. C. Representative western blot demonstrating that protein reduction due to siRNA to eIF4E or Ezrin occurs in invaded cells, thus invaded cells are not a result of rescue from the siRNA. Results are similar to MM6 cells grown in suspension . D. Visualization of masks generated during Imaris confocal analysis to automate identification of pseudopods and subsequent measurement of contents. Scale bar = 10µm. E. Total mRNA (left) and rRNA (right) levels detected by RT-qPCR in invaded MM6 cells as a function of genetic knockdown using siRNA to EIF4E or EZR as compared to siLUC treated cells. Each symbol represents a biological replicate performed independently. Bars represent the mean, shown with standard deviations and p-values (Welch’s t test). F. Representative western blots of invaded CRISPR MM6 cells showing reduced levels of factors in the Ezrin-CD44-HA axis similar to cells grown in suspension.

Journal: bioRxiv

Article Title: eIF4E and Ezrin cooperate in pseudopods to drive a localized migratory translation program in acute myeloid leukemia

doi: 10.64898/2026.02.21.707190

Figure Lengend Snippet: A. Schema of VISTA-R for invaded cells. Steps for isolated invaded AML cells for marking active ribosomes with the VISTA-R method. B. Confocal microscopy for patient specimen, AML23, demonstrated active translation, characterized by VISTA-R and Ezrin are present in the same pseudopods (white arrow). A single section through the plane of the cell is shown. Scale bar = 10µm. C. Representative western blot demonstrating that protein reduction due to siRNA to eIF4E or Ezrin occurs in invaded cells, thus invaded cells are not a result of rescue from the siRNA. Results are similar to MM6 cells grown in suspension . D. Visualization of masks generated during Imaris confocal analysis to automate identification of pseudopods and subsequent measurement of contents. Scale bar = 10µm. E. Total mRNA (left) and rRNA (right) levels detected by RT-qPCR in invaded MM6 cells as a function of genetic knockdown using siRNA to EIF4E or EZR as compared to siLUC treated cells. Each symbol represents a biological replicate performed independently. Bars represent the mean, shown with standard deviations and p-values (Welch’s t test). F. Representative western blots of invaded CRISPR MM6 cells showing reduced levels of factors in the Ezrin-CD44-HA axis similar to cells grown in suspension.

Article Snippet: Antibodies for immunoblotting: mouse monoclonal anti-eIF4E (cat# 610270, BD Biosciences), mouse monoclonal anti-β-Actin (cat# A5441, Sigma Aldrich), rabbit polyclonal anti-Mcl-1 (S-19) (cat# sc-819, Santa Cruz), mouse monoclonal anti-HSP90α/β (F-8) (cat# sc-13119, Santa Cruz), rabbit polyclonal anti-Myc (cat# ab32072, Abcam), Mouse monoclonal anti-CD44 antibody (cat# 156–3 C11, Cell Signaling Technology), rabbit polyclonal anti-CD44 (cat# A12410, Abclonal), rabbit polyclonal anti-HAS3 antibody (cat# ab154104, Abcam), rabbit polyclonal anti-phosphoglucomutase 5 (cat# AI14638, Abgent), rabbit polyclonal anti-Lamin A (C-terminal) (cat# L1293, Sigma Aldrich), rabbit polyclonal anti-GAPDH (FL-335) (cat# sc-25778, Santa Cruz), rabbit polyclonal anti-UGDH (cat#AP12613b-EV, Abgent), rabbit monoclonal anti-Hexokinase II (cat# 2867, Cell Signaling Technology), rabbit monoclonal anti-UAP1 antibody (cat# 2716, GenuinBiotech), mouse monoclonal anti-RPL17 (C-8, cat# sc-515904, Santa Cruz), mouse monoclonal anti-RPS16 (D-8, cat# sc-518206; Santa Cruz), mouse monoclonal anti-RPS6 (C-8, cat# sc-74459, Santa Cruz), mouse monoclonal anti-eIF3e (G-7, cat# sc-390413, Santa Cruz), mouse monoclonal anti-eIF3p110 (B-6, cat# sc-74507, Santa Cruz), mouse monoclonal anti-eIF3b (A-7, cat# sc-374156, Santa Cruz), mouse monoclonal anti-eIF4AI/II (H-5, cat# sc-377315, Santa Cruz), mouse monoclonal anti-eIF4G (A-10, cat# sc-133155, Santa Cruz), mouse monoclonal anti-Nopp140 (E-7, cat# sc-374033, Santa Cruz), rabbit monoclonal anti-histone H3 acetyl K27 (cat# ab177178, Abcam), rabbit polyclonal Calreticulin (cat# AW5211, ABCEPTA), rabbit polyclonal anti-Calnexin (cat# ab22595, Abcam), rabbit polyclonal anti-Ezrin antibody (cat# PA5-80603, Invitrogen), mouse monoclonal anti-Ezrin (CPTC-Ezrin-1, cat# AB_2100318, DSHB), mouse monoclonal anti-MEK-1 (H-8, sc-6250, Santa Cruz), and mouse monoclonal anti-cytochrome c (A-8, cat# sc-13156, Santa Cruz).

Techniques: Isolation, Confocal Microscopy, Western Blot, Suspension, Generated, Quantitative RT-PCR, Knockdown, CRISPR

A. Ezrin and eIF4E RIPs from cytoplasmic fractions from invaded MM6 cells compared to input. Each symbol represents a biological replicate. Means, standard deviations, p-values (Welch’s t test) are shown. Mitochondrial encoded mRNAs were used as negative controls (MT-CYB and MT-CO1). B. Representative western blots showing the expression for the indicated proteins using total cell lysates of invaded MM6 cells with knockdown of Ezrin (siEZR) or eIF4E (siEIF4E) and siLUC control. Quantitation of these western blots shown in C , each symbol represents a biological replicate, bar shows the mean, standard deviations and p-values (multiple unpaired t tests). D. Translation inhibitors decrease the adhesion capacity of MM6 cells. Each dot represents a biological replicate. Means, standard deviations and p-values (one way ANOVA) are shown.

Journal: bioRxiv

Article Title: eIF4E and Ezrin cooperate in pseudopods to drive a localized migratory translation program in acute myeloid leukemia

doi: 10.64898/2026.02.21.707190

Figure Lengend Snippet: A. Ezrin and eIF4E RIPs from cytoplasmic fractions from invaded MM6 cells compared to input. Each symbol represents a biological replicate. Means, standard deviations, p-values (Welch’s t test) are shown. Mitochondrial encoded mRNAs were used as negative controls (MT-CYB and MT-CO1). B. Representative western blots showing the expression for the indicated proteins using total cell lysates of invaded MM6 cells with knockdown of Ezrin (siEZR) or eIF4E (siEIF4E) and siLUC control. Quantitation of these western blots shown in C , each symbol represents a biological replicate, bar shows the mean, standard deviations and p-values (multiple unpaired t tests). D. Translation inhibitors decrease the adhesion capacity of MM6 cells. Each dot represents a biological replicate. Means, standard deviations and p-values (one way ANOVA) are shown.

Article Snippet: Antibodies for immunoblotting: mouse monoclonal anti-eIF4E (cat# 610270, BD Biosciences), mouse monoclonal anti-β-Actin (cat# A5441, Sigma Aldrich), rabbit polyclonal anti-Mcl-1 (S-19) (cat# sc-819, Santa Cruz), mouse monoclonal anti-HSP90α/β (F-8) (cat# sc-13119, Santa Cruz), rabbit polyclonal anti-Myc (cat# ab32072, Abcam), Mouse monoclonal anti-CD44 antibody (cat# 156–3 C11, Cell Signaling Technology), rabbit polyclonal anti-CD44 (cat# A12410, Abclonal), rabbit polyclonal anti-HAS3 antibody (cat# ab154104, Abcam), rabbit polyclonal anti-phosphoglucomutase 5 (cat# AI14638, Abgent), rabbit polyclonal anti-Lamin A (C-terminal) (cat# L1293, Sigma Aldrich), rabbit polyclonal anti-GAPDH (FL-335) (cat# sc-25778, Santa Cruz), rabbit polyclonal anti-UGDH (cat#AP12613b-EV, Abgent), rabbit monoclonal anti-Hexokinase II (cat# 2867, Cell Signaling Technology), rabbit monoclonal anti-UAP1 antibody (cat# 2716, GenuinBiotech), mouse monoclonal anti-RPL17 (C-8, cat# sc-515904, Santa Cruz), mouse monoclonal anti-RPS16 (D-8, cat# sc-518206; Santa Cruz), mouse monoclonal anti-RPS6 (C-8, cat# sc-74459, Santa Cruz), mouse monoclonal anti-eIF3e (G-7, cat# sc-390413, Santa Cruz), mouse monoclonal anti-eIF3p110 (B-6, cat# sc-74507, Santa Cruz), mouse monoclonal anti-eIF3b (A-7, cat# sc-374156, Santa Cruz), mouse monoclonal anti-eIF4AI/II (H-5, cat# sc-377315, Santa Cruz), mouse monoclonal anti-eIF4G (A-10, cat# sc-133155, Santa Cruz), mouse monoclonal anti-Nopp140 (E-7, cat# sc-374033, Santa Cruz), rabbit monoclonal anti-histone H3 acetyl K27 (cat# ab177178, Abcam), rabbit polyclonal Calreticulin (cat# AW5211, ABCEPTA), rabbit polyclonal anti-Calnexin (cat# ab22595, Abcam), rabbit polyclonal anti-Ezrin antibody (cat# PA5-80603, Invitrogen), mouse monoclonal anti-Ezrin (CPTC-Ezrin-1, cat# AB_2100318, DSHB), mouse monoclonal anti-MEK-1 (H-8, sc-6250, Santa Cruz), and mouse monoclonal anti-cytochrome c (A-8, cat# sc-13156, Santa Cruz).

Techniques: Western Blot, Expressing, Knockdown, Control, Quantitation Assay

A. Representative western blot from total cell lysates of MM6 and NOMO-1 cell lines demonstrating NOMO-1 cells have similar eIF4E levels to CD34+ from healthy donors, while MM6 cells have similar levels to high-eIF4E AML patient specimens. b-Actin is provided for loading control. Each lane refers to a different sample. B . Confocal micrograph of eIF4E nuclear and cytoplasmic staining in MM6 cells. Scale bar = 10 µm. C, D. Quantitation of western blots for NOMO-1 eIF4E relative to vector or MM6 CRISPR 4E relative to CRISPR-CTRL cells ( , Supplemental Figure 1G). Each data point represents a biological replicate. Bar represents the mean. Standard deviation and p-values (Welch’s t test) are shown. E. Representative confocal micrograph in NOMO-1 eIF4E cells demonstrating that HA staining is specific as its signal is removed upon treatment with hyaluronidase (HAse). HA is red; DAPI is blue. F. Representative western blot of MM6 cells treated with the eIF4E inhibitor ribavirin or vehicle control demonstrating that ribavirin reduces eIF4E target proteins including Ezrin. b-Actin is provided as a loading control. G, H . Representative western blots demonstrating lower eIF4E levels and factors in the Ezrin-CD44-HA axis in MM6 and THP-1 CRISPR-4E cells compared to CRISPR-Controls.

Journal: bioRxiv

Article Title: eIF4E and Ezrin cooperate in pseudopods to drive a localized migratory translation program in acute myeloid leukemia

doi: 10.64898/2026.02.21.707190

Figure Lengend Snippet: A. Representative western blot from total cell lysates of MM6 and NOMO-1 cell lines demonstrating NOMO-1 cells have similar eIF4E levels to CD34+ from healthy donors, while MM6 cells have similar levels to high-eIF4E AML patient specimens. b-Actin is provided for loading control. Each lane refers to a different sample. B . Confocal micrograph of eIF4E nuclear and cytoplasmic staining in MM6 cells. Scale bar = 10 µm. C, D. Quantitation of western blots for NOMO-1 eIF4E relative to vector or MM6 CRISPR 4E relative to CRISPR-CTRL cells ( , Supplemental Figure 1G). Each data point represents a biological replicate. Bar represents the mean. Standard deviation and p-values (Welch’s t test) are shown. E. Representative confocal micrograph in NOMO-1 eIF4E cells demonstrating that HA staining is specific as its signal is removed upon treatment with hyaluronidase (HAse). HA is red; DAPI is blue. F. Representative western blot of MM6 cells treated with the eIF4E inhibitor ribavirin or vehicle control demonstrating that ribavirin reduces eIF4E target proteins including Ezrin. b-Actin is provided as a loading control. G, H . Representative western blots demonstrating lower eIF4E levels and factors in the Ezrin-CD44-HA axis in MM6 and THP-1 CRISPR-4E cells compared to CRISPR-Controls.

Article Snippet: Antibodies for immunoblotting: mouse monoclonal anti-eIF4E (cat# 610270, BD Biosciences), mouse monoclonal anti-β-Actin (cat# A5441, Sigma Aldrich), rabbit polyclonal anti-Mcl-1 (S-19) (cat# sc-819, Santa Cruz), mouse monoclonal anti-HSP90α/β (F-8) (cat# sc-13119, Santa Cruz), rabbit polyclonal anti-Myc (cat# ab32072, Abcam), Mouse monoclonal anti-CD44 antibody (cat# 156–3 C11, Cell Signaling Technology), rabbit polyclonal anti-CD44 (cat# A12410, Abclonal), rabbit polyclonal anti-HAS3 antibody (cat# ab154104, Abcam), rabbit polyclonal anti-phosphoglucomutase 5 (cat# AI14638, Abgent), rabbit polyclonal anti-Lamin A (C-terminal) (cat# L1293, Sigma Aldrich), rabbit polyclonal anti-GAPDH (FL-335) (cat# sc-25778, Santa Cruz), rabbit polyclonal anti-UGDH (cat#AP12613b-EV, Abgent), rabbit monoclonal anti-Hexokinase II (cat# 2867, Cell Signaling Technology), rabbit monoclonal anti-UAP1 antibody (cat# 2716, GenuinBiotech), mouse monoclonal anti-RPL17 (C-8, cat# sc-515904, Santa Cruz), mouse monoclonal anti-RPS16 (D-8, cat# sc-518206; Santa Cruz), mouse monoclonal anti-RPS6 (C-8, cat# sc-74459, Santa Cruz), mouse monoclonal anti-eIF3e (G-7, cat# sc-390413, Santa Cruz), mouse monoclonal anti-eIF3p110 (B-6, cat# sc-74507, Santa Cruz), mouse monoclonal anti-eIF3b (A-7, cat# sc-374156, Santa Cruz), mouse monoclonal anti-eIF4AI/II (H-5, cat# sc-377315, Santa Cruz), mouse monoclonal anti-eIF4G (A-10, cat# sc-133155, Santa Cruz), mouse monoclonal anti-Nopp140 (E-7, cat# sc-374033, Santa Cruz), rabbit monoclonal anti-histone H3 acetyl K27 (cat# ab177178, Abcam), rabbit polyclonal Calreticulin (cat# AW5211, ABCEPTA), rabbit polyclonal anti-Calnexin (cat# ab22595, Abcam), rabbit polyclonal anti-Ezrin antibody (cat# PA5-80603, Invitrogen), mouse monoclonal anti-Ezrin (CPTC-Ezrin-1, cat# AB_2100318, DSHB), mouse monoclonal anti-MEK-1 (H-8, sc-6250, Santa Cruz), and mouse monoclonal anti-cytochrome c (A-8, cat# sc-13156, Santa Cruz).

Techniques: Western Blot, Control, Staining, Quantitation Assay, Plasmid Preparation, CRISPR, Standard Deviation

B. The impact on the Ezrin-CD44-HA axis of NOMO-1 eIF4E overexpression (eIF4E) compared to vector control (A) or of MM6 CRISPR-4E cells relative to MM6 CRISPR-CTRL cells (B). Left panels. Western blots of eIF4E and Ezrin-CD44-HA axis. Corresponding b-Actin shown for loading controls. Right panels . Confocal micrographs (single section through the plane of cells) of the indicated cells stained for HA or CD44 shown in red. DAPI shown in blue. Scale bar = 10µm. C. Western blot of primary specimens from AML patients (with normal or high-eIF4E) and CD34 + from healthy donors showing expression of eIF4E and Ezrin. b-Actin is provided as a loading control. Each lane represents a different individual. D. Adhesion (left) and invasion (right) capacity using HS-5 stromal cells of NOMO-1 and MM6 cell lines as a function of genetic eIF4E overexpression or CRISPR knockdown respectively. Data is presented as fold change relative to their corresponding controls. Each symbol represents an independent experiment performed with replicates. The bar represents the mean with standard deviations and p-values calculated with two tailed paired T test. E. Schematic representation of colonization assay into mesenchymal stromal cell spheroid model mimicking the bone marrow niche. F. Quantification of AML cell colonization capacity relative to their corresponding controls. Each symbol represents an experimental replicate. The bar represents the mean with standard deviations and p-values, Welch’s t test.

Journal: bioRxiv

Article Title: eIF4E and Ezrin cooperate in pseudopods to drive a localized migratory translation program in acute myeloid leukemia

doi: 10.64898/2026.02.21.707190

Figure Lengend Snippet: B. The impact on the Ezrin-CD44-HA axis of NOMO-1 eIF4E overexpression (eIF4E) compared to vector control (A) or of MM6 CRISPR-4E cells relative to MM6 CRISPR-CTRL cells (B). Left panels. Western blots of eIF4E and Ezrin-CD44-HA axis. Corresponding b-Actin shown for loading controls. Right panels . Confocal micrographs (single section through the plane of cells) of the indicated cells stained for HA or CD44 shown in red. DAPI shown in blue. Scale bar = 10µm. C. Western blot of primary specimens from AML patients (with normal or high-eIF4E) and CD34 + from healthy donors showing expression of eIF4E and Ezrin. b-Actin is provided as a loading control. Each lane represents a different individual. D. Adhesion (left) and invasion (right) capacity using HS-5 stromal cells of NOMO-1 and MM6 cell lines as a function of genetic eIF4E overexpression or CRISPR knockdown respectively. Data is presented as fold change relative to their corresponding controls. Each symbol represents an independent experiment performed with replicates. The bar represents the mean with standard deviations and p-values calculated with two tailed paired T test. E. Schematic representation of colonization assay into mesenchymal stromal cell spheroid model mimicking the bone marrow niche. F. Quantification of AML cell colonization capacity relative to their corresponding controls. Each symbol represents an experimental replicate. The bar represents the mean with standard deviations and p-values, Welch’s t test.

Article Snippet: Antibodies for immunoblotting: mouse monoclonal anti-eIF4E (cat# 610270, BD Biosciences), mouse monoclonal anti-β-Actin (cat# A5441, Sigma Aldrich), rabbit polyclonal anti-Mcl-1 (S-19) (cat# sc-819, Santa Cruz), mouse monoclonal anti-HSP90α/β (F-8) (cat# sc-13119, Santa Cruz), rabbit polyclonal anti-Myc (cat# ab32072, Abcam), Mouse monoclonal anti-CD44 antibody (cat# 156–3 C11, Cell Signaling Technology), rabbit polyclonal anti-CD44 (cat# A12410, Abclonal), rabbit polyclonal anti-HAS3 antibody (cat# ab154104, Abcam), rabbit polyclonal anti-phosphoglucomutase 5 (cat# AI14638, Abgent), rabbit polyclonal anti-Lamin A (C-terminal) (cat# L1293, Sigma Aldrich), rabbit polyclonal anti-GAPDH (FL-335) (cat# sc-25778, Santa Cruz), rabbit polyclonal anti-UGDH (cat#AP12613b-EV, Abgent), rabbit monoclonal anti-Hexokinase II (cat# 2867, Cell Signaling Technology), rabbit monoclonal anti-UAP1 antibody (cat# 2716, GenuinBiotech), mouse monoclonal anti-RPL17 (C-8, cat# sc-515904, Santa Cruz), mouse monoclonal anti-RPS16 (D-8, cat# sc-518206; Santa Cruz), mouse monoclonal anti-RPS6 (C-8, cat# sc-74459, Santa Cruz), mouse monoclonal anti-eIF3e (G-7, cat# sc-390413, Santa Cruz), mouse monoclonal anti-eIF3p110 (B-6, cat# sc-74507, Santa Cruz), mouse monoclonal anti-eIF3b (A-7, cat# sc-374156, Santa Cruz), mouse monoclonal anti-eIF4AI/II (H-5, cat# sc-377315, Santa Cruz), mouse monoclonal anti-eIF4G (A-10, cat# sc-133155, Santa Cruz), mouse monoclonal anti-Nopp140 (E-7, cat# sc-374033, Santa Cruz), rabbit monoclonal anti-histone H3 acetyl K27 (cat# ab177178, Abcam), rabbit polyclonal Calreticulin (cat# AW5211, ABCEPTA), rabbit polyclonal anti-Calnexin (cat# ab22595, Abcam), rabbit polyclonal anti-Ezrin antibody (cat# PA5-80603, Invitrogen), mouse monoclonal anti-Ezrin (CPTC-Ezrin-1, cat# AB_2100318, DSHB), mouse monoclonal anti-MEK-1 (H-8, sc-6250, Santa Cruz), and mouse monoclonal anti-cytochrome c (A-8, cat# sc-13156, Santa Cruz).

Techniques: Over Expression, Plasmid Preparation, Control, CRISPR, Western Blot, Staining, Expressing, Knockdown, Two Tailed Test

A. Western blot of total cell lysates from MM6 cells grown in suspension demonstrated knockdown of Ezrin (siEZR) or eIF4E (siEIF4E) compared to RNAi to luciferase (siLUC) used as a negative control. b-Actin is provided as a loading control. Other proteins of the Ezrin-CD44-HA axis are also shown. Quantification for these is shown in with 3-6 biological replicates for each protein. B. Adhesion and invasion capacity of MM6 cells onto/through HS-5 bone marrow stroma. Fold change relative to siLUC is shown. Each symbol represents a biological replicate performed independently with replicates. Bars represent the mean, shown with standard deviations and p-values (one-way ANOVA). C. Western blot of eIF4E and Ezrin immunoprecipitations (IPs) using total cell lysates from MM6 cells in suspension. SN, supernatant after immunoprecipitation, IgG, negative control. Representative of three biological replicates. IPs of total cell lysates from THP-1 cells in suspension are provided in . D. IPs from MM6 cells in suspension using the rRNA antibody Y10b. LC indicates antibody light chain. Representative of three biological replicates. E. RNA immunoprecipitations (RIPs) from MM6 total cell lysates grown in suspension using anti-Ezrin (Ezrin RIP) or anti-eIF4E (eIF4E RIP) antibodies. Data are from RT-qPCR and represented relative to input. Each symbol represents a biological replicate performed independently with triplicates. Bars represent the mean, shown with standard deviations and p-values (two tailed Welch’s t test). F. Western blots of eIF4E and Ezrin IPs from the cytoplasmic fractions of MM6 cells in suspension or G . After invasion through HS-5 bone marrow stroma (invaded). Fractionation controls are provided in . H. RIPs from MM6 cytoplasmic fraction from invaded cells using anti-eIF4E (eIF4E RIP) or anti-Ezrin (Ezrin RIP) antibodies. Data are from RT-qPCR represented relative to input. Each symbol represents a biological replicate performed independently with triplicates. Bars represent the mean, shown with standard deviations and p-values (two tailed Welch’s t test). I. Count of the number of pseudopods observed in suspension and invaded MM6 cells represented as a fraction relative to the total cells counted. Each symbol represents a biological replicate. Bars represent the mean, shown with standard deviations and p-values (two-way ANOVA). J . Immunofluorescence and confocal microscopy demonstrating eIF4E, Ezrin, CD44 and rRNA are localized to the same pseudopods (white arrows). All confocal micrographs represent a single section through the plane of the cell. Scale bar = 10 µm

Journal: bioRxiv

Article Title: eIF4E and Ezrin cooperate in pseudopods to drive a localized migratory translation program in acute myeloid leukemia

doi: 10.64898/2026.02.21.707190

Figure Lengend Snippet: A. Western blot of total cell lysates from MM6 cells grown in suspension demonstrated knockdown of Ezrin (siEZR) or eIF4E (siEIF4E) compared to RNAi to luciferase (siLUC) used as a negative control. b-Actin is provided as a loading control. Other proteins of the Ezrin-CD44-HA axis are also shown. Quantification for these is shown in with 3-6 biological replicates for each protein. B. Adhesion and invasion capacity of MM6 cells onto/through HS-5 bone marrow stroma. Fold change relative to siLUC is shown. Each symbol represents a biological replicate performed independently with replicates. Bars represent the mean, shown with standard deviations and p-values (one-way ANOVA). C. Western blot of eIF4E and Ezrin immunoprecipitations (IPs) using total cell lysates from MM6 cells in suspension. SN, supernatant after immunoprecipitation, IgG, negative control. Representative of three biological replicates. IPs of total cell lysates from THP-1 cells in suspension are provided in . D. IPs from MM6 cells in suspension using the rRNA antibody Y10b. LC indicates antibody light chain. Representative of three biological replicates. E. RNA immunoprecipitations (RIPs) from MM6 total cell lysates grown in suspension using anti-Ezrin (Ezrin RIP) or anti-eIF4E (eIF4E RIP) antibodies. Data are from RT-qPCR and represented relative to input. Each symbol represents a biological replicate performed independently with triplicates. Bars represent the mean, shown with standard deviations and p-values (two tailed Welch’s t test). F. Western blots of eIF4E and Ezrin IPs from the cytoplasmic fractions of MM6 cells in suspension or G . After invasion through HS-5 bone marrow stroma (invaded). Fractionation controls are provided in . H. RIPs from MM6 cytoplasmic fraction from invaded cells using anti-eIF4E (eIF4E RIP) or anti-Ezrin (Ezrin RIP) antibodies. Data are from RT-qPCR represented relative to input. Each symbol represents a biological replicate performed independently with triplicates. Bars represent the mean, shown with standard deviations and p-values (two tailed Welch’s t test). I. Count of the number of pseudopods observed in suspension and invaded MM6 cells represented as a fraction relative to the total cells counted. Each symbol represents a biological replicate. Bars represent the mean, shown with standard deviations and p-values (two-way ANOVA). J . Immunofluorescence and confocal microscopy demonstrating eIF4E, Ezrin, CD44 and rRNA are localized to the same pseudopods (white arrows). All confocal micrographs represent a single section through the plane of the cell. Scale bar = 10 µm

Article Snippet: Antibodies for immunoblotting: mouse monoclonal anti-eIF4E (cat# 610270, BD Biosciences), mouse monoclonal anti-β-Actin (cat# A5441, Sigma Aldrich), rabbit polyclonal anti-Mcl-1 (S-19) (cat# sc-819, Santa Cruz), mouse monoclonal anti-HSP90α/β (F-8) (cat# sc-13119, Santa Cruz), rabbit polyclonal anti-Myc (cat# ab32072, Abcam), Mouse monoclonal anti-CD44 antibody (cat# 156–3 C11, Cell Signaling Technology), rabbit polyclonal anti-CD44 (cat# A12410, Abclonal), rabbit polyclonal anti-HAS3 antibody (cat# ab154104, Abcam), rabbit polyclonal anti-phosphoglucomutase 5 (cat# AI14638, Abgent), rabbit polyclonal anti-Lamin A (C-terminal) (cat# L1293, Sigma Aldrich), rabbit polyclonal anti-GAPDH (FL-335) (cat# sc-25778, Santa Cruz), rabbit polyclonal anti-UGDH (cat#AP12613b-EV, Abgent), rabbit monoclonal anti-Hexokinase II (cat# 2867, Cell Signaling Technology), rabbit monoclonal anti-UAP1 antibody (cat# 2716, GenuinBiotech), mouse monoclonal anti-RPL17 (C-8, cat# sc-515904, Santa Cruz), mouse monoclonal anti-RPS16 (D-8, cat# sc-518206; Santa Cruz), mouse monoclonal anti-RPS6 (C-8, cat# sc-74459, Santa Cruz), mouse monoclonal anti-eIF3e (G-7, cat# sc-390413, Santa Cruz), mouse monoclonal anti-eIF3p110 (B-6, cat# sc-74507, Santa Cruz), mouse monoclonal anti-eIF3b (A-7, cat# sc-374156, Santa Cruz), mouse monoclonal anti-eIF4AI/II (H-5, cat# sc-377315, Santa Cruz), mouse monoclonal anti-eIF4G (A-10, cat# sc-133155, Santa Cruz), mouse monoclonal anti-Nopp140 (E-7, cat# sc-374033, Santa Cruz), rabbit monoclonal anti-histone H3 acetyl K27 (cat# ab177178, Abcam), rabbit polyclonal Calreticulin (cat# AW5211, ABCEPTA), rabbit polyclonal anti-Calnexin (cat# ab22595, Abcam), rabbit polyclonal anti-Ezrin antibody (cat# PA5-80603, Invitrogen), mouse monoclonal anti-Ezrin (CPTC-Ezrin-1, cat# AB_2100318, DSHB), mouse monoclonal anti-MEK-1 (H-8, sc-6250, Santa Cruz), and mouse monoclonal anti-cytochrome c (A-8, cat# sc-13156, Santa Cruz).

Techniques: Western Blot, Suspension, Knockdown, Luciferase, Negative Control, Control, Immunoprecipitation, Quantitative RT-PCR, Two Tailed Test, Fractionation, Immunofluorescence, Confocal Microscopy

A. Genetic reduction of Ezrin and eIF4E using siRNA in MM6 cells grown in suspension impacts production of factors in the Ezrin-CD44-HA axis compared to the siLUC control shown in a representative western blot (left panel). b-Actin is provided as loading control. Right panel, quantification of protein expression for the indicated siRNAs relative to siLUC is shown. The expression of each protein was calculated relative to b-Actin. Each symbol represents a biological replicate. Means, standard deviations and p-values (multiple paired t tests). B. Endogenous Ezrin and eIF4E immunoprecipitations (IP) in THP-1 total cell lysates show similar observations to MM6 cells . SN supernatant after immunoprecipitation, IgG negative control. H2B also serves as a negative control for eIF4E and Ezrin IPs. C. Cytoplasmic (left) and total (right) cell lysates demonstrate Ezrin immunoprecipitated with the translation machinery including eIF4AI/II. D. Example fractionation controls for suspension (left) or invaded (right) cells shown in indicating quality of fractions. MEK and CalR are cytoplasmic markers; NOPP140 and H3K27A are nuclear markers.

Journal: bioRxiv

Article Title: eIF4E and Ezrin cooperate in pseudopods to drive a localized migratory translation program in acute myeloid leukemia

doi: 10.64898/2026.02.21.707190

Figure Lengend Snippet: A. Genetic reduction of Ezrin and eIF4E using siRNA in MM6 cells grown in suspension impacts production of factors in the Ezrin-CD44-HA axis compared to the siLUC control shown in a representative western blot (left panel). b-Actin is provided as loading control. Right panel, quantification of protein expression for the indicated siRNAs relative to siLUC is shown. The expression of each protein was calculated relative to b-Actin. Each symbol represents a biological replicate. Means, standard deviations and p-values (multiple paired t tests). B. Endogenous Ezrin and eIF4E immunoprecipitations (IP) in THP-1 total cell lysates show similar observations to MM6 cells . SN supernatant after immunoprecipitation, IgG negative control. H2B also serves as a negative control for eIF4E and Ezrin IPs. C. Cytoplasmic (left) and total (right) cell lysates demonstrate Ezrin immunoprecipitated with the translation machinery including eIF4AI/II. D. Example fractionation controls for suspension (left) or invaded (right) cells shown in indicating quality of fractions. MEK and CalR are cytoplasmic markers; NOPP140 and H3K27A are nuclear markers.

Article Snippet: Antibodies for immunoblotting: mouse monoclonal anti-eIF4E (cat# 610270, BD Biosciences), mouse monoclonal anti-β-Actin (cat# A5441, Sigma Aldrich), rabbit polyclonal anti-Mcl-1 (S-19) (cat# sc-819, Santa Cruz), mouse monoclonal anti-HSP90α/β (F-8) (cat# sc-13119, Santa Cruz), rabbit polyclonal anti-Myc (cat# ab32072, Abcam), Mouse monoclonal anti-CD44 antibody (cat# 156–3 C11, Cell Signaling Technology), rabbit polyclonal anti-CD44 (cat# A12410, Abclonal), rabbit polyclonal anti-HAS3 antibody (cat# ab154104, Abcam), rabbit polyclonal anti-phosphoglucomutase 5 (cat# AI14638, Abgent), rabbit polyclonal anti-Lamin A (C-terminal) (cat# L1293, Sigma Aldrich), rabbit polyclonal anti-GAPDH (FL-335) (cat# sc-25778, Santa Cruz), rabbit polyclonal anti-UGDH (cat#AP12613b-EV, Abgent), rabbit monoclonal anti-Hexokinase II (cat# 2867, Cell Signaling Technology), rabbit monoclonal anti-UAP1 antibody (cat# 2716, GenuinBiotech), mouse monoclonal anti-RPL17 (C-8, cat# sc-515904, Santa Cruz), mouse monoclonal anti-RPS16 (D-8, cat# sc-518206; Santa Cruz), mouse monoclonal anti-RPS6 (C-8, cat# sc-74459, Santa Cruz), mouse monoclonal anti-eIF3e (G-7, cat# sc-390413, Santa Cruz), mouse monoclonal anti-eIF3p110 (B-6, cat# sc-74507, Santa Cruz), mouse monoclonal anti-eIF3b (A-7, cat# sc-374156, Santa Cruz), mouse monoclonal anti-eIF4AI/II (H-5, cat# sc-377315, Santa Cruz), mouse monoclonal anti-eIF4G (A-10, cat# sc-133155, Santa Cruz), mouse monoclonal anti-Nopp140 (E-7, cat# sc-374033, Santa Cruz), rabbit monoclonal anti-histone H3 acetyl K27 (cat# ab177178, Abcam), rabbit polyclonal Calreticulin (cat# AW5211, ABCEPTA), rabbit polyclonal anti-Calnexin (cat# ab22595, Abcam), rabbit polyclonal anti-Ezrin antibody (cat# PA5-80603, Invitrogen), mouse monoclonal anti-Ezrin (CPTC-Ezrin-1, cat# AB_2100318, DSHB), mouse monoclonal anti-MEK-1 (H-8, sc-6250, Santa Cruz), and mouse monoclonal anti-cytochrome c (A-8, cat# sc-13156, Santa Cruz).

Techniques: Suspension, Control, Western Blot, Expressing, Immunoprecipitation, Negative Control, Fractionation

A. Western blots of total cell lysates for MM6 cells in suspension treated with puromycin and analyzed by WB with anti-puromycin antibody (to assess global protein synthesis) reveal that Ezrin does not influence global translation: RNAi knockdown to Ezrin (siEZR) did not show difference compared to luciferase (siLUC) control (left panel). In contrast, CRISPR 4E reduces global translation signal by about 50% relative to CRISPR-CTRL, consistent with previous studies. Each symbol represents a biological replicate, bar showing mean, standard deviation, p-value (two tailed paired t test). B. Confocal micrographs for VISTA-R and the indicated proteins for MM6 cells upon genetic knockdowns siEIF4E), siEZR or siLUC control Arrows indicate T-PODs. Scale bar = 10µm. Western blots confirmed knockdown (Supplementary Figure 6C) C. As in B but monitoring eIF4E in T-PODs as a function of Ezrin and eIF4E knockdown. Ezrin is also reduced by eIF4E knockdown in invaded cells (Supplementary Figure 6C). Scale bar = 10µm. D. Quantitation of intensity of factors as noted in T-PODs over 3 biological replicates for VISTA-R and rRNA, 5 for Ezrin and 2 for eIF4E. Boxplots show Imaris-quantified TPOD-associated intensities of VISTA-R, rRNA, Ezrin, and eIF4E under the conditions shown. Each dot represents an individual T-POD object, p-values calculated using one way ANOVA test.

Journal: bioRxiv

Article Title: eIF4E and Ezrin cooperate in pseudopods to drive a localized migratory translation program in acute myeloid leukemia

doi: 10.64898/2026.02.21.707190

Figure Lengend Snippet: A. Western blots of total cell lysates for MM6 cells in suspension treated with puromycin and analyzed by WB with anti-puromycin antibody (to assess global protein synthesis) reveal that Ezrin does not influence global translation: RNAi knockdown to Ezrin (siEZR) did not show difference compared to luciferase (siLUC) control (left panel). In contrast, CRISPR 4E reduces global translation signal by about 50% relative to CRISPR-CTRL, consistent with previous studies. Each symbol represents a biological replicate, bar showing mean, standard deviation, p-value (two tailed paired t test). B. Confocal micrographs for VISTA-R and the indicated proteins for MM6 cells upon genetic knockdowns siEIF4E), siEZR or siLUC control Arrows indicate T-PODs. Scale bar = 10µm. Western blots confirmed knockdown (Supplementary Figure 6C) C. As in B but monitoring eIF4E in T-PODs as a function of Ezrin and eIF4E knockdown. Ezrin is also reduced by eIF4E knockdown in invaded cells (Supplementary Figure 6C). Scale bar = 10µm. D. Quantitation of intensity of factors as noted in T-PODs over 3 biological replicates for VISTA-R and rRNA, 5 for Ezrin and 2 for eIF4E. Boxplots show Imaris-quantified TPOD-associated intensities of VISTA-R, rRNA, Ezrin, and eIF4E under the conditions shown. Each dot represents an individual T-POD object, p-values calculated using one way ANOVA test.

Article Snippet: Antibodies for immunoblotting: mouse monoclonal anti-eIF4E (cat# 610270, BD Biosciences), mouse monoclonal anti-β-Actin (cat# A5441, Sigma Aldrich), rabbit polyclonal anti-Mcl-1 (S-19) (cat# sc-819, Santa Cruz), mouse monoclonal anti-HSP90α/β (F-8) (cat# sc-13119, Santa Cruz), rabbit polyclonal anti-Myc (cat# ab32072, Abcam), Mouse monoclonal anti-CD44 antibody (cat# 156–3 C11, Cell Signaling Technology), rabbit polyclonal anti-CD44 (cat# A12410, Abclonal), rabbit polyclonal anti-HAS3 antibody (cat# ab154104, Abcam), rabbit polyclonal anti-phosphoglucomutase 5 (cat# AI14638, Abgent), rabbit polyclonal anti-Lamin A (C-terminal) (cat# L1293, Sigma Aldrich), rabbit polyclonal anti-GAPDH (FL-335) (cat# sc-25778, Santa Cruz), rabbit polyclonal anti-UGDH (cat#AP12613b-EV, Abgent), rabbit monoclonal anti-Hexokinase II (cat# 2867, Cell Signaling Technology), rabbit monoclonal anti-UAP1 antibody (cat# 2716, GenuinBiotech), mouse monoclonal anti-RPL17 (C-8, cat# sc-515904, Santa Cruz), mouse monoclonal anti-RPS16 (D-8, cat# sc-518206; Santa Cruz), mouse monoclonal anti-RPS6 (C-8, cat# sc-74459, Santa Cruz), mouse monoclonal anti-eIF3e (G-7, cat# sc-390413, Santa Cruz), mouse monoclonal anti-eIF3p110 (B-6, cat# sc-74507, Santa Cruz), mouse monoclonal anti-eIF3b (A-7, cat# sc-374156, Santa Cruz), mouse monoclonal anti-eIF4AI/II (H-5, cat# sc-377315, Santa Cruz), mouse monoclonal anti-eIF4G (A-10, cat# sc-133155, Santa Cruz), mouse monoclonal anti-Nopp140 (E-7, cat# sc-374033, Santa Cruz), rabbit monoclonal anti-histone H3 acetyl K27 (cat# ab177178, Abcam), rabbit polyclonal Calreticulin (cat# AW5211, ABCEPTA), rabbit polyclonal anti-Calnexin (cat# ab22595, Abcam), rabbit polyclonal anti-Ezrin antibody (cat# PA5-80603, Invitrogen), mouse monoclonal anti-Ezrin (CPTC-Ezrin-1, cat# AB_2100318, DSHB), mouse monoclonal anti-MEK-1 (H-8, sc-6250, Santa Cruz), and mouse monoclonal anti-cytochrome c (A-8, cat# sc-13156, Santa Cruz).

Techniques: Western Blot, Suspension, Knockdown, Luciferase, Control, CRISPR, Standard Deviation, Two Tailed Test, Quantitation Assay

A . Schematic comparing previous strategies to monitor in situ translation based on surrogate markers of translation such as labelled amino acids (left), and our novel method for direct measurement of translation, VISTA-R (right). B. VISTA-R signals in MM6 cells are directly related to translation activity since these are abrogated upon addition of translation inhibitors. Puromycin and/or HHT were added prior to the fixation step in VISTA-R. Representative confocal micrograph of VISTA-R signal in vehicle control, puromycin, HHT or combination treated cells. DAPI is shown in blue. Scale bar = 10 µm. C. Quantitation showing means, standard deviations of the mean, data points from each measurement, p-values calculated with one way ANOVA test from over 60 cells. D. T-PODs (as shown by arrows) contain active ribosomes as seen by VISTA-R (green). VISTA-R and rRNA (red), Ezrin (blue) overlay in these T-POD. E. Same as D but demonstrating VISTA-R (green), Ezrin (magenta) and RPS6 (red) and CD44 (turquoise) are present in the same T-PODs (see arrows). F. Same as D-E but demonstrating that VISTA-R (green), eIF4E (red) and Ezrin (magenta) and CD44 (turquoise) are present in the same T-PODs (see arrows). All confocal micrographs represent a single section through the plane of the cell. Scale bars = 10 µm.

Journal: bioRxiv

Article Title: eIF4E and Ezrin cooperate in pseudopods to drive a localized migratory translation program in acute myeloid leukemia

doi: 10.64898/2026.02.21.707190

Figure Lengend Snippet: A . Schematic comparing previous strategies to monitor in situ translation based on surrogate markers of translation such as labelled amino acids (left), and our novel method for direct measurement of translation, VISTA-R (right). B. VISTA-R signals in MM6 cells are directly related to translation activity since these are abrogated upon addition of translation inhibitors. Puromycin and/or HHT were added prior to the fixation step in VISTA-R. Representative confocal micrograph of VISTA-R signal in vehicle control, puromycin, HHT or combination treated cells. DAPI is shown in blue. Scale bar = 10 µm. C. Quantitation showing means, standard deviations of the mean, data points from each measurement, p-values calculated with one way ANOVA test from over 60 cells. D. T-PODs (as shown by arrows) contain active ribosomes as seen by VISTA-R (green). VISTA-R and rRNA (red), Ezrin (blue) overlay in these T-POD. E. Same as D but demonstrating VISTA-R (green), Ezrin (magenta) and RPS6 (red) and CD44 (turquoise) are present in the same T-PODs (see arrows). F. Same as D-E but demonstrating that VISTA-R (green), eIF4E (red) and Ezrin (magenta) and CD44 (turquoise) are present in the same T-PODs (see arrows). All confocal micrographs represent a single section through the plane of the cell. Scale bars = 10 µm.

Article Snippet: Antibodies for immunoblotting: mouse monoclonal anti-eIF4E (cat# 610270, BD Biosciences), mouse monoclonal anti-β-Actin (cat# A5441, Sigma Aldrich), rabbit polyclonal anti-Mcl-1 (S-19) (cat# sc-819, Santa Cruz), mouse monoclonal anti-HSP90α/β (F-8) (cat# sc-13119, Santa Cruz), rabbit polyclonal anti-Myc (cat# ab32072, Abcam), Mouse monoclonal anti-CD44 antibody (cat# 156–3 C11, Cell Signaling Technology), rabbit polyclonal anti-CD44 (cat# A12410, Abclonal), rabbit polyclonal anti-HAS3 antibody (cat# ab154104, Abcam), rabbit polyclonal anti-phosphoglucomutase 5 (cat# AI14638, Abgent), rabbit polyclonal anti-Lamin A (C-terminal) (cat# L1293, Sigma Aldrich), rabbit polyclonal anti-GAPDH (FL-335) (cat# sc-25778, Santa Cruz), rabbit polyclonal anti-UGDH (cat#AP12613b-EV, Abgent), rabbit monoclonal anti-Hexokinase II (cat# 2867, Cell Signaling Technology), rabbit monoclonal anti-UAP1 antibody (cat# 2716, GenuinBiotech), mouse monoclonal anti-RPL17 (C-8, cat# sc-515904, Santa Cruz), mouse monoclonal anti-RPS16 (D-8, cat# sc-518206; Santa Cruz), mouse monoclonal anti-RPS6 (C-8, cat# sc-74459, Santa Cruz), mouse monoclonal anti-eIF3e (G-7, cat# sc-390413, Santa Cruz), mouse monoclonal anti-eIF3p110 (B-6, cat# sc-74507, Santa Cruz), mouse monoclonal anti-eIF3b (A-7, cat# sc-374156, Santa Cruz), mouse monoclonal anti-eIF4AI/II (H-5, cat# sc-377315, Santa Cruz), mouse monoclonal anti-eIF4G (A-10, cat# sc-133155, Santa Cruz), mouse monoclonal anti-Nopp140 (E-7, cat# sc-374033, Santa Cruz), rabbit monoclonal anti-histone H3 acetyl K27 (cat# ab177178, Abcam), rabbit polyclonal Calreticulin (cat# AW5211, ABCEPTA), rabbit polyclonal anti-Calnexin (cat# ab22595, Abcam), rabbit polyclonal anti-Ezrin antibody (cat# PA5-80603, Invitrogen), mouse monoclonal anti-Ezrin (CPTC-Ezrin-1, cat# AB_2100318, DSHB), mouse monoclonal anti-MEK-1 (H-8, sc-6250, Santa Cruz), and mouse monoclonal anti-cytochrome c (A-8, cat# sc-13156, Santa Cruz).

Techniques: In Situ, Activity Assay, Control, Quantitation Assay

A. Schema of VISTA-R for invaded cells. Steps for isolated invaded AML cells for marking active ribosomes with the VISTA-R method. B. Confocal microscopy for patient specimen, AML23, demonstrated active translation, characterized by VISTA-R and Ezrin are present in the same pseudopods (white arrow). A single section through the plane of the cell is shown. Scale bar = 10µm. C. Representative western blot demonstrating that protein reduction due to siRNA to eIF4E or Ezrin occurs in invaded cells, thus invaded cells are not a result of rescue from the siRNA. Results are similar to MM6 cells grown in suspension . D. Visualization of masks generated during Imaris confocal analysis to automate identification of pseudopods and subsequent measurement of contents. Scale bar = 10µm. E. Total mRNA (left) and rRNA (right) levels detected by RT-qPCR in invaded MM6 cells as a function of genetic knockdown using siRNA to EIF4E or EZR as compared to siLUC treated cells. Each symbol represents a biological replicate performed independently. Bars represent the mean, shown with standard deviations and p-values (Welch’s t test). F. Representative western blots of invaded CRISPR MM6 cells showing reduced levels of factors in the Ezrin-CD44-HA axis similar to cells grown in suspension.

Journal: bioRxiv

Article Title: eIF4E and Ezrin cooperate in pseudopods to drive a localized migratory translation program in acute myeloid leukemia

doi: 10.64898/2026.02.21.707190

Figure Lengend Snippet: A. Schema of VISTA-R for invaded cells. Steps for isolated invaded AML cells for marking active ribosomes with the VISTA-R method. B. Confocal microscopy for patient specimen, AML23, demonstrated active translation, characterized by VISTA-R and Ezrin are present in the same pseudopods (white arrow). A single section through the plane of the cell is shown. Scale bar = 10µm. C. Representative western blot demonstrating that protein reduction due to siRNA to eIF4E or Ezrin occurs in invaded cells, thus invaded cells are not a result of rescue from the siRNA. Results are similar to MM6 cells grown in suspension . D. Visualization of masks generated during Imaris confocal analysis to automate identification of pseudopods and subsequent measurement of contents. Scale bar = 10µm. E. Total mRNA (left) and rRNA (right) levels detected by RT-qPCR in invaded MM6 cells as a function of genetic knockdown using siRNA to EIF4E or EZR as compared to siLUC treated cells. Each symbol represents a biological replicate performed independently. Bars represent the mean, shown with standard deviations and p-values (Welch’s t test). F. Representative western blots of invaded CRISPR MM6 cells showing reduced levels of factors in the Ezrin-CD44-HA axis similar to cells grown in suspension.

Article Snippet: Antibodies for immunoblotting: mouse monoclonal anti-eIF4E (cat# 610270, BD Biosciences), mouse monoclonal anti-β-Actin (cat# A5441, Sigma Aldrich), rabbit polyclonal anti-Mcl-1 (S-19) (cat# sc-819, Santa Cruz), mouse monoclonal anti-HSP90α/β (F-8) (cat# sc-13119, Santa Cruz), rabbit polyclonal anti-Myc (cat# ab32072, Abcam), Mouse monoclonal anti-CD44 antibody (cat# 156–3 C11, Cell Signaling Technology), rabbit polyclonal anti-CD44 (cat# A12410, Abclonal), rabbit polyclonal anti-HAS3 antibody (cat# ab154104, Abcam), rabbit polyclonal anti-phosphoglucomutase 5 (cat# AI14638, Abgent), rabbit polyclonal anti-Lamin A (C-terminal) (cat# L1293, Sigma Aldrich), rabbit polyclonal anti-GAPDH (FL-335) (cat# sc-25778, Santa Cruz), rabbit polyclonal anti-UGDH (cat#AP12613b-EV, Abgent), rabbit monoclonal anti-Hexokinase II (cat# 2867, Cell Signaling Technology), rabbit monoclonal anti-UAP1 antibody (cat# 2716, GenuinBiotech), mouse monoclonal anti-RPL17 (C-8, cat# sc-515904, Santa Cruz), mouse monoclonal anti-RPS16 (D-8, cat# sc-518206; Santa Cruz), mouse monoclonal anti-RPS6 (C-8, cat# sc-74459, Santa Cruz), mouse monoclonal anti-eIF3e (G-7, cat# sc-390413, Santa Cruz), mouse monoclonal anti-eIF3p110 (B-6, cat# sc-74507, Santa Cruz), mouse monoclonal anti-eIF3b (A-7, cat# sc-374156, Santa Cruz), mouse monoclonal anti-eIF4AI/II (H-5, cat# sc-377315, Santa Cruz), mouse monoclonal anti-eIF4G (A-10, cat# sc-133155, Santa Cruz), mouse monoclonal anti-Nopp140 (E-7, cat# sc-374033, Santa Cruz), rabbit monoclonal anti-histone H3 acetyl K27 (cat# ab177178, Abcam), rabbit polyclonal Calreticulin (cat# AW5211, ABCEPTA), rabbit polyclonal anti-Calnexin (cat# ab22595, Abcam), rabbit polyclonal anti-Ezrin antibody (cat# PA5-80603, Invitrogen), mouse monoclonal anti-Ezrin (CPTC-Ezrin-1, cat# AB_2100318, DSHB), mouse monoclonal anti-MEK-1 (H-8, sc-6250, Santa Cruz), and mouse monoclonal anti-cytochrome c (A-8, cat# sc-13156, Santa Cruz).

Techniques: Isolation, Confocal Microscopy, Western Blot, Suspension, Generated, Quantitative RT-PCR, Knockdown, CRISPR

A. Ezrin and eIF4E RIPs from cytoplasmic fractions from invaded MM6 cells compared to input. Each symbol represents a biological replicate. Means, standard deviations, p-values (Welch’s t test) are shown. Mitochondrial encoded mRNAs were used as negative controls (MT-CYB and MT-CO1). B. Representative western blots showing the expression for the indicated proteins using total cell lysates of invaded MM6 cells with knockdown of Ezrin (siEZR) or eIF4E (siEIF4E) and siLUC control. Quantitation of these western blots shown in C , each symbol represents a biological replicate, bar shows the mean, standard deviations and p-values (multiple unpaired t tests). D. Translation inhibitors decrease the adhesion capacity of MM6 cells. Each dot represents a biological replicate. Means, standard deviations and p-values (one way ANOVA) are shown.

Journal: bioRxiv

Article Title: eIF4E and Ezrin cooperate in pseudopods to drive a localized migratory translation program in acute myeloid leukemia

doi: 10.64898/2026.02.21.707190

Figure Lengend Snippet: A. Ezrin and eIF4E RIPs from cytoplasmic fractions from invaded MM6 cells compared to input. Each symbol represents a biological replicate. Means, standard deviations, p-values (Welch’s t test) are shown. Mitochondrial encoded mRNAs were used as negative controls (MT-CYB and MT-CO1). B. Representative western blots showing the expression for the indicated proteins using total cell lysates of invaded MM6 cells with knockdown of Ezrin (siEZR) or eIF4E (siEIF4E) and siLUC control. Quantitation of these western blots shown in C , each symbol represents a biological replicate, bar shows the mean, standard deviations and p-values (multiple unpaired t tests). D. Translation inhibitors decrease the adhesion capacity of MM6 cells. Each dot represents a biological replicate. Means, standard deviations and p-values (one way ANOVA) are shown.

Article Snippet: Antibodies for immunoblotting: mouse monoclonal anti-eIF4E (cat# 610270, BD Biosciences), mouse monoclonal anti-β-Actin (cat# A5441, Sigma Aldrich), rabbit polyclonal anti-Mcl-1 (S-19) (cat# sc-819, Santa Cruz), mouse monoclonal anti-HSP90α/β (F-8) (cat# sc-13119, Santa Cruz), rabbit polyclonal anti-Myc (cat# ab32072, Abcam), Mouse monoclonal anti-CD44 antibody (cat# 156–3 C11, Cell Signaling Technology), rabbit polyclonal anti-CD44 (cat# A12410, Abclonal), rabbit polyclonal anti-HAS3 antibody (cat# ab154104, Abcam), rabbit polyclonal anti-phosphoglucomutase 5 (cat# AI14638, Abgent), rabbit polyclonal anti-Lamin A (C-terminal) (cat# L1293, Sigma Aldrich), rabbit polyclonal anti-GAPDH (FL-335) (cat# sc-25778, Santa Cruz), rabbit polyclonal anti-UGDH (cat#AP12613b-EV, Abgent), rabbit monoclonal anti-Hexokinase II (cat# 2867, Cell Signaling Technology), rabbit monoclonal anti-UAP1 antibody (cat# 2716, GenuinBiotech), mouse monoclonal anti-RPL17 (C-8, cat# sc-515904, Santa Cruz), mouse monoclonal anti-RPS16 (D-8, cat# sc-518206; Santa Cruz), mouse monoclonal anti-RPS6 (C-8, cat# sc-74459, Santa Cruz), mouse monoclonal anti-eIF3e (G-7, cat# sc-390413, Santa Cruz), mouse monoclonal anti-eIF3p110 (B-6, cat# sc-74507, Santa Cruz), mouse monoclonal anti-eIF3b (A-7, cat# sc-374156, Santa Cruz), mouse monoclonal anti-eIF4AI/II (H-5, cat# sc-377315, Santa Cruz), mouse monoclonal anti-eIF4G (A-10, cat# sc-133155, Santa Cruz), mouse monoclonal anti-Nopp140 (E-7, cat# sc-374033, Santa Cruz), rabbit monoclonal anti-histone H3 acetyl K27 (cat# ab177178, Abcam), rabbit polyclonal Calreticulin (cat# AW5211, ABCEPTA), rabbit polyclonal anti-Calnexin (cat# ab22595, Abcam), rabbit polyclonal anti-Ezrin antibody (cat# PA5-80603, Invitrogen), mouse monoclonal anti-Ezrin (CPTC-Ezrin-1, cat# AB_2100318, DSHB), mouse monoclonal anti-MEK-1 (H-8, sc-6250, Santa Cruz), and mouse monoclonal anti-cytochrome c (A-8, cat# sc-13156, Santa Cruz).

Techniques: Western Blot, Expressing, Knockdown, Control, Quantitation Assay

Journal: iScience

Article Title: RhoA downregulation in the murine intestinal epithelium results in chronic Wnt activation and increased tumorigenesis

doi: 10.1016/j.isci.2024.109400

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-phospho-ERM (Ezrin, Thr567; Radixin, Thr564; Moesin, Thr558) , Cell Signaling Technology , Cat#3141; RRID: AB_330232.

Techniques: Recombinant, Polymer, Fluorescence, Reverse Transcription, Activation Assay, Software