antibody to eif4e  (Cell Signaling Technology Inc)

Journal: Cell Biology and Toxicology

Article Title: Circular RNA circTrpc6-induced tau phosphorylation contributes to neuronal injury and cognitive decline via miR-485-5p/eIF4E axis in senescence-accelerated mouse prone 8 mice

doi: 10.1007/s10565-026-10214-y

Figure Lengend Snippet: eIF4E is the downstream target of circTrpc6. (A) A volcano plot showing dysregulated proteins in three hippocampal samples from control and circTrpc6-overexpressing mice via proteomics, N = 3 samples/group. (B) Cytoscape software with Degree, MCC, and MNC algorithms was employed to identify the top ten hub proteins based on their ranking scores. (C) Relative expression of circTrpc6 was evaluated via RT-PCR assay after N2a cells was infected with LVs containing circTrpc6 shRNA, N = 3. (D) The effect of circTrpc6 knockdown on eIF4E and p-eIF4E expression in N2a cells was assessed using Western blot assay. N = 4. (E) The effect of circTrpc6 knockdown on eIF4E and p-eIF4E expression in the hippocampus of SAMP8 mice was assessed using Western blot assay. N = 4. (F) Relative expression of circTrpc6 was evaluated via RT-PCR assay after N2a cells was infected with circTrpc6 overexpression LVs, N = 3. (G) The effect of circTrpc6 overexpression on eIF4E and p-eIF4E expression in N2a cells was assessed using Western blot assay. N = 4. All data in the figure are presented as mean ± SEM. Student’s t test was used to assess statistically significant differences. ** P < 0.01

Article Snippet: In brief, cells were collected and lysed, and the resultant lysates were incubated with protein A/G agarose beads (Santa Cruz Biotechnology, USA) along with an eIF4E antibody (ABclonal, China) overnight at 4 °C.

Techniques: Control, Software, Expressing, Reverse Transcription Polymerase Chain Reaction, Infection, shRNA, Knockdown, Western Blot, Over Expression

Journal: Cell Biology and Toxicology

Article Title: Circular RNA circTrpc6-induced tau phosphorylation contributes to neuronal injury and cognitive decline via miR-485-5p/eIF4E axis in senescence-accelerated mouse prone 8 mice

doi: 10.1007/s10565-026-10214-y

Figure Lengend Snippet: Effect of eIF4E knockdown on cognitive function, hippocampal neuronal and synaptic injury in SAMP8 mice. (A) Relative expression of eIF4E in 3-month-old SAMP8 and age-matched SAMR1 mice was detected via RT-PCR assay. N = 3 mice. (B) RT-PCR assay was used to detect eIF4E expression in the hippocampus after 3-month-old SAMP8 was infected with eIF4E shRNA AAVs for 6 weeks, N = 3 mice. (C ~ F) The effect of eIF4E knockdown on the cognitive function of SAMP8 mice was evaluated via the MWM test ( C ), passive avoidance test ( D ), and novel object recognition task ( E, F ), N = 8 ~ 10 mice. (G ~ I) The effect of eIF4E knockdown on the percentage of Nissl body ( G ), neuronal apoptotic rate ( H ) and spine density ( I ) in the hippocampus of SAMP8 mice were evaluated via Nissl, TUNEL and Golgi staining assays, respectively, N = 3 mice. All data in the figure are presented as mean ± SEM. Student’s t test was used to assess statistically significant differences. * P < 0.05, ** P < 0.01

Article Snippet: In brief, cells were collected and lysed, and the resultant lysates were incubated with protein A/G agarose beads (Santa Cruz Biotechnology, USA) along with an eIF4E antibody (ABclonal, China) overnight at 4 °C.

Techniques: Knockdown, Expressing, Reverse Transcription Polymerase Chain Reaction, Infection, shRNA, TUNEL Assay, Staining

Journal: Cell Biology and Toxicology

Article Title: Circular RNA circTrpc6-induced tau phosphorylation contributes to neuronal injury and cognitive decline via miR-485-5p/eIF4E axis in senescence-accelerated mouse prone 8 mice

doi: 10.1007/s10565-026-10214-y

Figure Lengend Snippet: CircTrpc6 regulates tau phosphorylation via eIF4E. (A) The effect of eIF4E knockdown on the expression of eIF4E, p-eIF4E, tau and p-tau (S396) in N2a cells was detected via Western blot assay. N = 4. (B) The effect of eIF4E knockdown on tau and p-tau expression in the hippocampus of SAMP8 mice was assessed using Western blot assay. N = 4. (C) The effect of circTrpc6 knockdown on tau and p-tau (S396) expression in N2a cells was assessed using Western blot assay. N = 4. (D) The effect of circTrpc6 knockdown on tau and p-tau (S396) expression in the hippocampus of SAMP8 mice was assessed using Western blot assay. N = 4. (E) The effect of eIF4E overexpression on the expression of eIF4E, p-eIF4E, tau and p-tau (S396) in N2a cells was detected via Western blot assay. N = 4. (F) The effect of circTrpc6 overexpression on tau and p-tau (S396) expression in N2a cells was detected via Western blot assay. N = 4. (G) The effect of eIF4E knockdown on tau and p-tau (S396) expression regulated by circTrpc6 overexpression in N2a cells was detected via Western blot assay. N = 4. (H, I) The effect of circTrpc6 and eIF4E knockdown on tau aggregates in the hippocampus of SAMP8 mice was evaluated via ThS staining assay. N = 3 mice. (J, K) The effect of circTrpc6 and eIF4E overexpression on aggregates in N2a cells was evaluated via ThS staining assay. N = 3. (L) The effect of eIF4E knockdown on aggregates regulated by circTrpc6 overexpression in N2a cells was detected via ThS staining assay. N = 3. All data in the figure are presented as mean ± SEM. Student’s t test ( A ~ F, H ~ K ) and one-way ANOVA ( G, L ) were used to assess statistically significant differences. * P < 0.05, ** P < 0.01, ns, not significant

Article Snippet: In brief, cells were collected and lysed, and the resultant lysates were incubated with protein A/G agarose beads (Santa Cruz Biotechnology, USA) along with an eIF4E antibody (ABclonal, China) overnight at 4 °C.

Techniques: Phospho-proteomics, Knockdown, Expressing, Western Blot, Over Expression, Staining

Journal: Cell Biology and Toxicology

Article Title: Circular RNA circTrpc6-induced tau phosphorylation contributes to neuronal injury and cognitive decline via miR-485-5p/eIF4E axis in senescence-accelerated mouse prone 8 mice

doi: 10.1007/s10565-026-10214-y

Figure Lengend Snippet: CircTrpc6 regulates eIF4E expression via miR-485-5p. (A) Prediction of miRNAs interacting with circTrpc6 using miRanda and RNAhybrid and with eIF4E mRNA 3’UTR using TargetScan and miRDB. (B, C) The interactions of miR-485-5p between circTrpc6 (B) and eIF4E mRNA 3’UTR (C) were assessed via RNA pull-down assay, N = 3. (D, E) The effect of circTrpc6 knockdown on miR-485-5p expression in N2a cells (D) and the hippocampus of SAMP8 mice (E) was evaluated via RT-PCR assay. N = 3. (F) The effect of circTrpc6 overexpression on miR-485-5p expression in N2a cells was assessed via RT-PCR assay. N = 3. (G, H) The effect of miR-485-5p overexpression on eIF4E and p-eIF4E expression in N2a cells (G) and the hippocampus of SAMP8 mice (H) was detected via Western blot assay. N = 4. (I) The effect of miR-485-5p knockdown on eIF4E and p-eIF4E expression in N2a cells was detected via Western blot assay. N = 4. (J) The effect of miR-485-5p overexpression on eIF4E and p-eIF4E expression regulated by circTrpc6 overexpression in N2a cells was detected via Western blot assay. N = 4. All data in the figure are presented as mean ± SEM. Student’s t test ( B ~ I ) and one-way ANOVA ( J ) were used to assess statistically significant differences. * P < 0.05, ** P < 0.01

Article Snippet: In brief, cells were collected and lysed, and the resultant lysates were incubated with protein A/G agarose beads (Santa Cruz Biotechnology, USA) along with an eIF4E antibody (ABclonal, China) overnight at 4 °C.

Techniques: Expressing, Pull Down Assay, Knockdown, Reverse Transcription Polymerase Chain Reaction, Over Expression, Western Blot

Journal: Cell Biology and Toxicology

Article Title: Circular RNA circTrpc6-induced tau phosphorylation contributes to neuronal injury and cognitive decline via miR-485-5p/eIF4E axis in senescence-accelerated mouse prone 8 mice

doi: 10.1007/s10565-026-10214-y

Figure Lengend Snippet: Effect of miR-485-5p overexpression on cognitive function, tau phosphorylation, hippocampal tau aggregates, neuronal and synaptic injury in SAMP8 mice, and on tau phosphorylation and aggregation in N2a cells. (A) Relative expression of miR-485-5p in 3-month-old SAMP8 and age-matched SAMR1 mice was detected via RT-PCR assay. N = 3 mice. (B ~ E) The effect of miR-485-5p overexpression on the cognitive function of SAMP8 mice was evaluated via the MWM test ( B ), passive avoidance test ( C ), and novel object recognition task ( D, E ), N = 8 ~ 10 mice. (F, G) The effect of miR-485-5p overexpression on tau and p-tau (S396) expression in N2a cells (F) and the hippocampus of SAMP8 mice (G) was detected via Western blot assay. N = 4. (H) The effect of miR-485-5p knockdown on tau and p-tau (S396) expression in N2a cells was detected via Western blot assay. N = 4. (I) The effect of miR-485-5p knockdown on aggregates in N2a cells was evaluated via ThS staining assay. N = 3. (J) The effect of miR-485-5p overexpression on tau aggregates in the hippocampus of SAMP8 mice was assessed via ThS staining assay. N = 3. (K ~ M) The effect of eIF4E knockdown on the percentage of Nissl body ( K ), neuronal apoptotic rate ( L ) and spine density ( M ) in the hippocampus of SAMP8 mice were evaluated via Nissl, TUNEL and Golgi staining assays, respectively, N = 3 mice. All data in the figure are presented as mean ± SEM. Student’s t test was used to assess statistically significant differences. * P < 0.05, ** P < 0.01, ns, not significant

Article Snippet: In brief, cells were collected and lysed, and the resultant lysates were incubated with protein A/G agarose beads (Santa Cruz Biotechnology, USA) along with an eIF4E antibody (ABclonal, China) overnight at 4 °C.

Techniques: Over Expression, Phospho-proteomics, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Knockdown, Staining, TUNEL Assay

Journal: Cell Biology and Toxicology

Article Title: Circular RNA circTrpc6-induced tau phosphorylation contributes to neuronal injury and cognitive decline via miR-485-5p/eIF4E axis in senescence-accelerated mouse prone 8 mice

doi: 10.1007/s10565-026-10214-y

Figure Lengend Snippet: CircTrpc6 regulates tau phosphorylation and the interaction of eIF4E with tau mRNA via miR-485-5p/eIF4E axis. (A) The effect of miR-485-5p overexpression on tau and p-tau (S396) expression regulated by circTrpc6 overexpression in N2a cells was detected via Western blot assay. N = 4. (B) The effect of miR-485-5p overexpression on aggregates regulated by circTrpc6 overexpression in N2a cells was assessed via ThS staining assay. N = 3. (C) The effect of eIF4E knockdown on tau and p-tau (S396) expression regulated by miR-485-5p knockdown in N2a cells was detected via Western blot assay. N = 4. (D) The effect of eIF4E knockdown on aggregates regulated by miR-485-5p knockdown in N2a cells was assessed via ThS staining assay. N = 3. (E ~ G) The effects of eIF4E (E) and circTrpc6 (F) overexpression and miR-485-5p knockdown (G) on the interaction of eIF4E with tau mRNA were evaluated via RIP assay. N = 3. (H) The effect of miR-485-5p overexpression on the interaction of eIF4E with tau mRNA regulated by circTrpc6 overexpression in N2a cells was detected via RIP assay. N = 3. (I) The effect of eIF4E knockdown on the interaction of eIF4E with tau mRNA regulated by circTrpc6 overexpression in N2a cells was detected via RIP assay. N = 3. (J) The effect of eIF4E knockdown on the interaction of eIF4E with tau mRNA regulated by miR-485-5p knockdown in N2a cells was detected via RIP assay. N = 3. All data in the figure are presented as mean ± SEM. Student’s t test ( E ~ G ) and one-way ANOVA ( A ~ D, H ~ J ). ** P < 0.01, ns, not significant

Article Snippet: In brief, cells were collected and lysed, and the resultant lysates were incubated with protein A/G agarose beads (Santa Cruz Biotechnology, USA) along with an eIF4E antibody (ABclonal, China) overnight at 4 °C.

Techniques: Phospho-proteomics, Over Expression, Expressing, Western Blot, Staining, Knockdown

Journal: bioRxiv

Article Title: The enteroviral protease target LSM14A operates outside of P-bodies to augment antiviral innate immunity

doi: 10.64898/2026.05.11.723630

Figure Lengend Snippet: A. 293T cells were either treated with sodium arsenite (NaAsO 2 ) (100 µM for 1h at 37°C) or DMSO (vehicle), followed by staining with anti-LSM14A antibody. The graph on the right shows the number of LSM14A+ puncta in 25 randomly selected cells. Each dot represents a cell. Statistical significance was determined using two-tailed non-parametric t -test. *P < 0.05. B. 293T cells were stained with anti-LSM14A and either anti-DDX6 antibodies (Top panel) or with anti-4E-T antibodies (Bottom panel) followed by confocal microscopy. The nuclei were stained with DAPI. C. 293T cells treated with either vehicle or NaAsO 2 as in (A) were stained with anti-LSM14A and anti-G3BP1 antibodies, followed by confocal microscopy. White arrows indicate PBs juxtaposed to SGs.

Article Snippet: The antibodies used in this study include rabbit anti-LSM14A (Bethyl Laboratories; #A305-103A), mouse anti-LSM14A (Santa Cruz Biotechnology; #sc-398633), anti-DDX6 (Bethyl Laboratories; #A300-461A), anti-4E-T (Bethyl Laboratories; #A300-706A), anti-PEX19 (Proteintech; #14713-1-AP), anti-G3BP1 (Santa Cruz Biotechnology; #sc-365338), anti-PMP70 (Sigma-Aldrich; #SAB4200181), anti-HA (ABclonal; #AE008), anti-GAPDH (GeneTex; #GTX627408), and anti-β-actin (Thermo Fisher Scientific; #AM4302).

Techniques: Staining, Two Tailed Test, Confocal Microscopy

Journal: iScience

Article Title: Alox15 via H 2 O 2 mediates TP receptor palmitoylation and its membrane trafficking leading to platelet activation

doi: 10.1016/j.isci.2026.114796

Figure Lengend Snippet: 12/15-LOX is required for TXA2-induced platelet activation and hemostasis (A–C) Eight-weeks-old WT and 12/15-LOX −/− mice were subjected to measurement of body weight (A), tail bleeding time (B), and whole blood clotting time (C) ( n = 10). (D) Platelet-rich plasma (PRP) from WT and 12/15-LOX −/− mice were incubated with and without F 2 -TXA2 (1 μM) for the indicated periods at RT and photographed. The percentage of clot retraction and extruded serum volume were calculated as described in the methods ( n = 3). (E) Wahed platelets were plated onto fibrinogen-coated coverslips and after 1 h stained with phalloidin and DAPI and observed under a Zeiss inverted microscope (Axiovision Observer.z1; 40×/NA 0.6). The pictures were captured by a Zeiss AxioCam MRm camera using the microscope operating and image analysis software ZEN 2.6. (F) Washed platelets from WT mice were labeled with calcein acetoxymethyl ester (10 μM) for 30 min and placed onto fibrinogen-coated wells in a 96-well plate. Platelets were then incubated with and without F 2 -TXA2 at the indicated concentrations for 30 min, washed with PBS and the bound platelets were lysed with lysis buffer and the fluorescence intensity was measured at 494 excitation and 517 emission ( n = 3). (G) PRP from WT mice treated with and without F 2 -TXA2 at the indicated concentrations was subjected to aggregation assay in an aggregometer ( n = 3). (H) Washed platelets from WT and 12/15-LOX −/− mice were subjected adhesion assay as shown in panel F ( n = 3). (I) PRP from WT and 12/15-LOX −/− mice with and without the indicated treatments were subjected to aggregation assay in an aggregometer ( n = 3). (J) Washed platelets from WT and 12/15-LOX −/− mice were incubated with and without F 2 -TXA2 for 30 min and plated onto fibrinogen-coated coverslips for 1 h. Platelets were then fixed, permeabilized, and stained with phalloidin to visualize F-actin, and pictures were captured. (K and L) Platelets from WT and 12/15-LOX −/− mice were incubated with and without F 2 -TXA2 (1 μM) for indicated time periods, and RNA and protein extracts were prepared and analyzed by qRT-PCR (K) and western blotting (L) for 12-LOX, 12/15-LOX and β-actin mRNA and protein levels using their specific primers or antibodies, respectively ( n = 3). (M) Platelets from WT and 12/15-LOX −/− mice were treated with and without F 2 -TXA2 for 30 min, and protein extracts were prepared and analyzed by western blotting for the levels of phospho and total eIF4E and 4EBP1 using their specific antibodies ( n = 3). (N) All the conditions were the same as in panel M except that the extracts were immunoprecipitated with anti-4EBP1 antibody, and the immunocomplexes were analyzed by western blotting for eIF4E and normalized for 4EBP1. The input protein was analyzed for β-actin levels. (O and P) Platelets from WT mice were incubated with and without F 2 -TXA2 in the presence and absence of rapamycin (100 nM) or torin1 (100 nM) for 30 min, and protein extracts were analyzed by western blotting for p4EBP1, 4EBP1, 12/15-LOX, and β-actin levels using their specific antibodies ( n = 3). (Q) Platelets from WT mice and 12/15-LOX −/− mice were assessed for 12(S)-HETE levels using a kit from Cayman ( n = 7). (R–W) Platelets from WT mice and 12/15-LOX −/− mice were treated with and without U46619 (1 μM) or ADP (40 μM) for 30 min and 12(S)-HETE levels were measured (R and U) ( n = 7) or subjected to adhesion assay (S and V) ( n = 3) or aggregation assay (T and W) ( n = 3). All data are presented as mean ± SD and analyzed by paired Student’s t test. ∗ p < 0.01 versus WT mice or control; # p < 0.01 versus F 2 -TXA2 or WT + F 2 -TXA2 or U46619. Scale bars: 10 μm in (E) and (J).

Article Snippet: Anti-eIF4E antibody , Santa Cruz Biotechnology , sc-9976.

Techniques: Activation Assay, Coagulation, Clinical Proteomics, Incubation, Staining, Inverted Microscopy, Microscopy, Software, Labeling, Lysis, Fluorescence, Cell Adhesion Assay, Quantitative RT-PCR, Western Blot, Immunoprecipitation, Control

Journal: iScience

Article Title: Alox15 via H 2 O 2 mediates TP receptor palmitoylation and its membrane trafficking leading to platelet activation

doi: 10.1016/j.isci.2026.114796

Figure Lengend Snippet: 12/15-LOX is required for TXA2-induced platelet activation and hemostasis (A–C) Eight-weeks-old WT and 12/15-LOX −/− mice were subjected to measurement of body weight (A), tail bleeding time (B), and whole blood clotting time (C) ( n = 10). (D) Platelet-rich plasma (PRP) from WT and 12/15-LOX −/− mice were incubated with and without F 2 -TXA2 (1 μM) for the indicated periods at RT and photographed. The percentage of clot retraction and extruded serum volume were calculated as described in the methods ( n = 3). (E) Wahed platelets were plated onto fibrinogen-coated coverslips and after 1 h stained with phalloidin and DAPI and observed under a Zeiss inverted microscope (Axiovision Observer.z1; 40×/NA 0.6). The pictures were captured by a Zeiss AxioCam MRm camera using the microscope operating and image analysis software ZEN 2.6. (F) Washed platelets from WT mice were labeled with calcein acetoxymethyl ester (10 μM) for 30 min and placed onto fibrinogen-coated wells in a 96-well plate. Platelets were then incubated with and without F 2 -TXA2 at the indicated concentrations for 30 min, washed with PBS and the bound platelets were lysed with lysis buffer and the fluorescence intensity was measured at 494 excitation and 517 emission ( n = 3). (G) PRP from WT mice treated with and without F 2 -TXA2 at the indicated concentrations was subjected to aggregation assay in an aggregometer ( n = 3). (H) Washed platelets from WT and 12/15-LOX −/− mice were subjected adhesion assay as shown in panel F ( n = 3). (I) PRP from WT and 12/15-LOX −/− mice with and without the indicated treatments were subjected to aggregation assay in an aggregometer ( n = 3). (J) Washed platelets from WT and 12/15-LOX −/− mice were incubated with and without F 2 -TXA2 for 30 min and plated onto fibrinogen-coated coverslips for 1 h. Platelets were then fixed, permeabilized, and stained with phalloidin to visualize F-actin, and pictures were captured. (K and L) Platelets from WT and 12/15-LOX −/− mice were incubated with and without F 2 -TXA2 (1 μM) for indicated time periods, and RNA and protein extracts were prepared and analyzed by qRT-PCR (K) and western blotting (L) for 12-LOX, 12/15-LOX and β-actin mRNA and protein levels using their specific primers or antibodies, respectively ( n = 3). (M) Platelets from WT and 12/15-LOX −/− mice were treated with and without F 2 -TXA2 for 30 min, and protein extracts were prepared and analyzed by western blotting for the levels of phospho and total eIF4E and 4EBP1 using their specific antibodies ( n = 3). (N) All the conditions were the same as in panel M except that the extracts were immunoprecipitated with anti-4EBP1 antibody, and the immunocomplexes were analyzed by western blotting for eIF4E and normalized for 4EBP1. The input protein was analyzed for β-actin levels. (O and P) Platelets from WT mice were incubated with and without F 2 -TXA2 in the presence and absence of rapamycin (100 nM) or torin1 (100 nM) for 30 min, and protein extracts were analyzed by western blotting for p4EBP1, 4EBP1, 12/15-LOX, and β-actin levels using their specific antibodies ( n = 3). (Q) Platelets from WT mice and 12/15-LOX −/− mice were assessed for 12(S)-HETE levels using a kit from Cayman ( n = 7). (R–W) Platelets from WT mice and 12/15-LOX −/− mice were treated with and without U46619 (1 μM) or ADP (40 μM) for 30 min and 12(S)-HETE levels were measured (R and U) ( n = 7) or subjected to adhesion assay (S and V) ( n = 3) or aggregation assay (T and W) ( n = 3). All data are presented as mean ± SD and analyzed by paired Student’s t test. ∗ p < 0.01 versus WT mice or control; # p < 0.01 versus F 2 -TXA2 or WT + F 2 -TXA2 or U46619. Scale bars: 10 μm in (E) and (J).

Article Snippet: Anti-phospho eIF4E antibody , Cell Signaling Technology , 9741.

Techniques: Activation Assay, Coagulation, Clinical Proteomics, Incubation, Staining, Inverted Microscopy, Microscopy, Software, Labeling, Lysis, Fluorescence, Cell Adhesion Assay, Quantitative RT-PCR, Western Blot, Immunoprecipitation, Control