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Bio-Rad rat anti mouse cd44 monoclonal antibody
Rat Anti Mouse Cd44 Monoclonal Antibody, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 93/100, based on 37 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/mouse+monoclonal+cd44/pm41610521-54-3-10?v=Bio-Rad
Average 93 stars, based on 37 article reviews

rat anti mouse cd44 monoclonal antibody - by Bioz Stars, 2026-07

93/100 stars

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Histopathological comparison of tumor architecture and proliferation between patient renal cell carcinoma (RCC) specimens and corresponding patient-derived xenograft models. Formalin-fixed, paraffin-embedded sections from primary patient tumors (KiCa-Pt58 and KiCa-Pt118), matched subcutaneous xenografts, and intra-renal patient-derived orthotopic xenograft (PDOX) tumors were analyzed to assess preservation of tumor histology and proliferative characteristics. ( A , C ) Representative images of KiCa-Pt58 ( A ) and KiCa-Pt118 ( C ) tissues stained with hematoxylin and eosin (H&E) for tissue architecture or were subjected to immunohistochemistry (IHC) for human CD44 (tumor cell marker) and human Ki67 (proliferation marker). Brown staining indicates positive immunoreactivity. Images were captured at 100× original magnification using an Axiovert 200M deconvolution microscope and SlideBook 6.0 software (Intelligent Imaging Innovations, Denver, CO, USA). ( B , D ) Quantitative analysis of Ki67-positive area (%) in KiCa-Pt58 ( B ) and KiCa-Pt118 ( D ) tissues. Positive (brown) staining areas were quantified digitally using Adobe Photoshop 7.0, with the percentage of immunoreactive area calculated per field. Data are presented as mean ± SEM (multiple fields per sample). Comparisons among patient biopsy specimens, subcutaneous xenografts, and orthotopic PDOX tumors were performed using unpaired Student’s t -test. No significant differences were observed (ns, p > 0.05), demonstrating faithful recapitulation of the parental tumor proliferative index across model passages. Abbreviations: RCC, renal cell carcinoma; PDOX, patient-derived orthotopic xenograft; H&E, hematoxylin and eosin; IHC, immunohistochemistry.

Journal: Cancers

Article Title: Optimizing Sequential Targeted Therapies in Advanced Renal Cell Carcinoma Using Patient-Derived Orthotopic Xenograft Mouse Avatars

doi: 10.3390/cancers18101615

Figure Lengend Snippet: Histopathological comparison of tumor architecture and proliferation between patient renal cell carcinoma (RCC) specimens and corresponding patient-derived xenograft models. Formalin-fixed, paraffin-embedded sections from primary patient tumors (KiCa-Pt58 and KiCa-Pt118), matched subcutaneous xenografts, and intra-renal patient-derived orthotopic xenograft (PDOX) tumors were analyzed to assess preservation of tumor histology and proliferative characteristics. ( A , C ) Representative images of KiCa-Pt58 ( A ) and KiCa-Pt118 ( C ) tissues stained with hematoxylin and eosin (H&E) for tissue architecture or were subjected to immunohistochemistry (IHC) for human CD44 (tumor cell marker) and human Ki67 (proliferation marker). Brown staining indicates positive immunoreactivity. Images were captured at 100× original magnification using an Axiovert 200M deconvolution microscope and SlideBook 6.0 software (Intelligent Imaging Innovations, Denver, CO, USA). ( B , D ) Quantitative analysis of Ki67-positive area (%) in KiCa-Pt58 ( B ) and KiCa-Pt118 ( D ) tissues. Positive (brown) staining areas were quantified digitally using Adobe Photoshop 7.0, with the percentage of immunoreactive area calculated per field. Data are presented as mean ± SEM (multiple fields per sample). Comparisons among patient biopsy specimens, subcutaneous xenografts, and orthotopic PDOX tumors were performed using unpaired Student’s t -test. No significant differences were observed (ns, p > 0.05), demonstrating faithful recapitulation of the parental tumor proliferative index across model passages. Abbreviations: RCC, renal cell carcinoma; PDOX, patient-derived orthotopic xenograft; H&E, hematoxylin and eosin; IHC, immunohistochemistry.

Article Snippet: Paraffin-embedded sections (5 μm) were stained with H&E or subjected to immunohistochemistry using primary antibodies against human Ki67 (proliferation marker; Thermo Fisher Scientific, Waltham, MA, USA; 1:200), human CD44 (tumor cell marker; Acris Antibodies, Rockville, MD, USA; 1:75), mouse CD31 (angiogenesis marker; Abcam, Cambridge, MA, USA; 1:200), and human PD-L1 (programmed death-ligand 1, immune checkpoint ligand; BioLegend, San Diego, CA, USA; 1:200) [ ].

Techniques: Comparison, Derivative Assay, Formalin-fixed Paraffin-Embedded, Preserving, Staining, Immunohistochemistry, Marker, Microscopy, Software, Imaging

Histopathological and immunohistochemical evaluation of targeted therapy responses in the KiCa-Pt58 patient-derived orthotopic xenograft (PDOX) model. ( A ) Representative images of left kidney tumors from KiCa-Pt58 PDOX mice after vehicle control or sequential targeted therapy (Everolimus→Sunitinib [E→S], Pazopanib→Sunitinib [P→S], Sunitinib→Everolimus [S→E], Pazopanib→Everolimus [P→E]). Formalin-fixed, paraffin-embedded sections were stained with hematoxylin and eosin (H&E) for tumor architecture or subjected to immunohistochemistry (IHC) for human CD44 (tumor cell marker), human Ki67 (proliferation marker), mouse CD31 (angiogenesis/endothelial marker), and human PD-L1 (immune checkpoint ligand). Brown staining indicates positive immunoreactivity. Images were acquired at 100× original magnification using an Axiovert 200M deconvolution microscope and SlideBook 6.0 software (Intelligent Imaging Innovations, Denver, CO, USA). ( B – D ) Quantitative analysis of positive staining area (%) for Ki67 ( B ), CD31 ( C ), and PD-L1 ( D ) across treatment groups. Positive (brown) areas were quantified digitally using Adobe Photoshop 7.0 (percentage immunoreactive area per field). Data are presented as mean ± SEM (multiple fields per sample; n = 7–9 mice per group). Statistical comparisons vs. control were performed using one-way ANOVA followed by Dunnett’s or Tukey’s post hoc tests (GraphPad Prism v7). Asterisks indicate significance: * p < 0.05; ** p < 0.01; *** p < 0.001. Effective regimens (particularly P→E and S→E) significantly reduced Ki67+ proliferation, CD31+ vascularity, and PD-L1 expression compared to control, consistent with antitumor and potential immunomodulatory effects. Abbreviations: PDOX, patient-derived orthotopic xenograft; IHC, immunohistochemistry; E, everolimus; S, sunitinib; P, pazopanib.

Journal: Cancers

Article Title: Optimizing Sequential Targeted Therapies in Advanced Renal Cell Carcinoma Using Patient-Derived Orthotopic Xenograft Mouse Avatars

doi: 10.3390/cancers18101615

Figure Lengend Snippet: Histopathological and immunohistochemical evaluation of targeted therapy responses in the KiCa-Pt58 patient-derived orthotopic xenograft (PDOX) model. ( A ) Representative images of left kidney tumors from KiCa-Pt58 PDOX mice after vehicle control or sequential targeted therapy (Everolimus→Sunitinib [E→S], Pazopanib→Sunitinib [P→S], Sunitinib→Everolimus [S→E], Pazopanib→Everolimus [P→E]). Formalin-fixed, paraffin-embedded sections were stained with hematoxylin and eosin (H&E) for tumor architecture or subjected to immunohistochemistry (IHC) for human CD44 (tumor cell marker), human Ki67 (proliferation marker), mouse CD31 (angiogenesis/endothelial marker), and human PD-L1 (immune checkpoint ligand). Brown staining indicates positive immunoreactivity. Images were acquired at 100× original magnification using an Axiovert 200M deconvolution microscope and SlideBook 6.0 software (Intelligent Imaging Innovations, Denver, CO, USA). ( B – D ) Quantitative analysis of positive staining area (%) for Ki67 ( B ), CD31 ( C ), and PD-L1 ( D ) across treatment groups. Positive (brown) areas were quantified digitally using Adobe Photoshop 7.0 (percentage immunoreactive area per field). Data are presented as mean ± SEM (multiple fields per sample; n = 7–9 mice per group). Statistical comparisons vs. control were performed using one-way ANOVA followed by Dunnett’s or Tukey’s post hoc tests (GraphPad Prism v7). Asterisks indicate significance: * p < 0.05; ** p < 0.01; *** p < 0.001. Effective regimens (particularly P→E and S→E) significantly reduced Ki67+ proliferation, CD31+ vascularity, and PD-L1 expression compared to control, consistent with antitumor and potential immunomodulatory effects. Abbreviations: PDOX, patient-derived orthotopic xenograft; IHC, immunohistochemistry; E, everolimus; S, sunitinib; P, pazopanib.

Techniques: Immunohistochemical staining, Derivative Assay, Control, Formalin-fixed Paraffin-Embedded, Staining, Immunohistochemistry, Marker, Microscopy, Software, Imaging, Expressing

Histopathological and immunohistochemical assessment of targeted therapy responses in the KiCa-Pt118 patient-derived orthotopic xenograft (PDOX) model. ( A ) Representative images of left kidney tumors from KiCa-Pt118 PDOX mice after vehicle control or sequential targeted therapy (Everolimus→Sunitinib [E→S], Pazopanib→Sunitinib [P→S], Sunitinib→Everolimus [S→E], Pazopanib→Everolimus [P→E]). Formalin-fixed, paraffin-embedded sections were stained with hematoxylin and eosin (H&E) for tumor architecture or subjected to immunohistochemistry (IHC) for human CD44 (tumor cell marker), human Ki67 (proliferation marker), mouse CD31 (angiogenesis/endothelial marker), and human PD-L1 (immune checkpoint ligand). Brown staining indicates positive immunoreactivity. Images were acquired at 100× original magnification using an Axiovert 200M deconvolution microscope and SlideBook 6.0 software (Intelligent Imaging Innovations, Denver, CO, USA). ( B – D ) Quantitative analysis of positive staining area (%) for Ki67 ( B ), CD31 ( C ), and PD-L1 ( D ) across treatment groups. Positive (brown) areas were quantified digitally using Adobe Photoshop 7.0 (percentage immunoreactive area per field), as described in . Data are presented as mean ± SEM (multiple fields per sample; n = 7–9 mice per group). Statistical comparisons vs. control were performed using one-way ANOVA followed by Dunnett’s or Tukey’s post hoc tests (GraphPad Prism v7). Asterisks indicate significance: * p < 0.05; ** p < 0.01; *** p < 0.001. Effective regimens (particularly S→E) significantly reduced Ki67+ proliferation, CD31+ vascularity, and PD-L1 expression compared to control, consistent with antitumor activity in this indolent, non-metastatic model. Abbreviations: PDOX, patient-derived orthotopic xenograft; IHC, immunohistochemistry; E, everolimus; S, sunitinib; P, pazopanib.

Journal: Cancers

Article Title: Optimizing Sequential Targeted Therapies in Advanced Renal Cell Carcinoma Using Patient-Derived Orthotopic Xenograft Mouse Avatars

doi: 10.3390/cancers18101615

Figure Lengend Snippet: Histopathological and immunohistochemical assessment of targeted therapy responses in the KiCa-Pt118 patient-derived orthotopic xenograft (PDOX) model. ( A ) Representative images of left kidney tumors from KiCa-Pt118 PDOX mice after vehicle control or sequential targeted therapy (Everolimus→Sunitinib [E→S], Pazopanib→Sunitinib [P→S], Sunitinib→Everolimus [S→E], Pazopanib→Everolimus [P→E]). Formalin-fixed, paraffin-embedded sections were stained with hematoxylin and eosin (H&E) for tumor architecture or subjected to immunohistochemistry (IHC) for human CD44 (tumor cell marker), human Ki67 (proliferation marker), mouse CD31 (angiogenesis/endothelial marker), and human PD-L1 (immune checkpoint ligand). Brown staining indicates positive immunoreactivity. Images were acquired at 100× original magnification using an Axiovert 200M deconvolution microscope and SlideBook 6.0 software (Intelligent Imaging Innovations, Denver, CO, USA). ( B – D ) Quantitative analysis of positive staining area (%) for Ki67 ( B ), CD31 ( C ), and PD-L1 ( D ) across treatment groups. Positive (brown) areas were quantified digitally using Adobe Photoshop 7.0 (percentage immunoreactive area per field), as described in . Data are presented as mean ± SEM (multiple fields per sample; n = 7–9 mice per group). Statistical comparisons vs. control were performed using one-way ANOVA followed by Dunnett’s or Tukey’s post hoc tests (GraphPad Prism v7). Asterisks indicate significance: * p < 0.05; ** p < 0.01; *** p < 0.001. Effective regimens (particularly S→E) significantly reduced Ki67+ proliferation, CD31+ vascularity, and PD-L1 expression compared to control, consistent with antitumor activity in this indolent, non-metastatic model. Abbreviations: PDOX, patient-derived orthotopic xenograft; IHC, immunohistochemistry; E, everolimus; S, sunitinib; P, pazopanib.

Techniques: Immunohistochemical staining, Derivative Assay, Control, Formalin-fixed Paraffin-Embedded, Staining, Immunohistochemistry, Marker, Microscopy, Software, Imaging, Expressing, Activity 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.

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

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.

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

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.

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.

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

(A) Volcano plot representation of global transcriptional changes in Control versus RGS11-OE MIA PaCa-2 as determined by RNA-seq analysis (n=3 samples/cell type). (B) Heatmap depicting top 20 up and downregulated genes (DEGs) in RGS11-OE MIA PaCa-2. The color gradient from red to green indicates log-transformed average fold-change values. (C) Immunoblots of RGS11-OE MIA PaCa-2 and RGS11-KD L3.6pl cells showing expression of stemness enriched proteins (CD24, CD44, CD133) when compared to their respective controls. β-actin is used as a loading control. Densitometric quantification using ImageJ is shown as ratio of protein-to-actin below each blot. (D) The enrichment plots of Gene Set Enrichment Analysis (GSEA) of sequencing results displaying the core enrichment of gene sets associated with myogenesis, hedgehog and kras signaling, EMT, apoptosis and apical Junction pathways in RGS11-OE MIA PaCa-2 cells based on normalized enrichment score (NES). (E) Graphical representation of pathways associated with enriched gene sets. The pathway network is generated using ShinyGO 0.80 after enriching for GO Biological Process with 0.05 FDR cutoff value. (F) Bar plot display (color based on adj p, length based on gene count) of top 20 terms over enriched in the RGS11-OE MIA PaCa-2 RNA Seq gene lists ran for GO Biological processes. The gene lists are filtered using adjusted p < 0.05. (G) Venn diagram depicting overlap between differentially expressed genes associated with top 4 pathways enriched in GO Biological Processes functional analysis.

Journal: Molecular cancer research : MCR

Article Title: Loss of Regulator of G protein signaling 11 promotes pro-tumorigenic features in pancreatic cancer

doi: 10.1158/1541-7786.MCR-25-0144

Figure Lengend Snippet: (A) Volcano plot representation of global transcriptional changes in Control versus RGS11-OE MIA PaCa-2 as determined by RNA-seq analysis (n=3 samples/cell type). (B) Heatmap depicting top 20 up and downregulated genes (DEGs) in RGS11-OE MIA PaCa-2. The color gradient from red to green indicates log-transformed average fold-change values. (C) Immunoblots of RGS11-OE MIA PaCa-2 and RGS11-KD L3.6pl cells showing expression of stemness enriched proteins (CD24, CD44, CD133) when compared to their respective controls. β-actin is used as a loading control. Densitometric quantification using ImageJ is shown as ratio of protein-to-actin below each blot. (D) The enrichment plots of Gene Set Enrichment Analysis (GSEA) of sequencing results displaying the core enrichment of gene sets associated with myogenesis, hedgehog and kras signaling, EMT, apoptosis and apical Junction pathways in RGS11-OE MIA PaCa-2 cells based on normalized enrichment score (NES). (E) Graphical representation of pathways associated with enriched gene sets. The pathway network is generated using ShinyGO 0.80 after enriching for GO Biological Process with 0.05 FDR cutoff value. (F) Bar plot display (color based on adj p, length based on gene count) of top 20 terms over enriched in the RGS11-OE MIA PaCa-2 RNA Seq gene lists ran for GO Biological processes. The gene lists are filtered using adjusted p < 0.05. (G) Venn diagram depicting overlap between differentially expressed genes associated with top 4 pathways enriched in GO Biological Processes functional analysis.

Article Snippet: For immunoblotting, rabbit polyclonal antibodies for E-cadherin (#3195, RRID:AB_2291471), N- cadherin (#13116, RRID:AB_2687616), Vimentin (#5741, RRID:AB_10695459), ZEB1 (#3396, RRID:AB_1904164), Caspase-3 (#14220, RRID:AB_2798429), PARP (#9532, RRID:AB_659884) Phospho-Stat3 (#9145S, RRID:AB_2491009), Stat3 (#9139S, RRID:AB_331757) and β-catenin (#8480S, RRID:AB_11127855), and mouse monoclonal antibody for CD44 (#3570S, RRID:AB_2076465), were purchased from Cell Signaling Technology (Danvers, MA).

Techniques: Control, RNA Sequencing, Transformation Assay, Western Blot, Expressing, Sequencing, Generated, Functional Assay

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