rabbit eif4e  (Novus Biologicals)

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: Selective targeting of type II tRNAs underlies SLFN14-mediated translational repression and its dysregulation by thrombocytopenia-linked mutations

doi: 10.64898/2026.02.03.703619

Figure Lengend Snippet: (A) 293T cells were induced to express SLFN14 for 6, 12, or 24 h, and cell viability was assessed using the MTT assay. (B) Western blot analysis of mCherry-SLFN14, HSP70, β-actin, and GAPDH expression. (C) Western blot analysis of mTORC1 pathway markers. (D) Western blot analysis of total eIF4E and phosphorylated eIF4E. (E) Schematic model illustrating how the IT-linked K219N mutation disrupts protein synthesis by shifting SLFN14 RNA cleavage activity toward type II tRNAs, leading to ribosome stalling, activation of compensatory stress pathways, and ultimately cell death.

Article Snippet: The antibodies against the following proteins were used in the immunoblotting: mCherry (Bio-techne, NBP2-25157), HSP70 (Santa Cruz, sc-137210), β-actin (Sigma, A2228), GAPDH (Cell Signaling Technology, #2118), 4E-BP1 (Cell Signaling Technology, #9452), Phospho-4E-BP1(Thr37/46) (Cell Signaling Technology, #2855), eIF4E (Cell Signaling Technology, #9742), Phospho-eIF4E (Ser209) (Cell Signaling Technology, #9741), p70 S6 Kinase (Cell Signaling Technology, #9202), Phospho-p70 S6 Kinase (Cell Signaling Technology, #9205).

Techniques: MTT Assay, Western Blot, Expressing, Mutagenesis, Activity Assay, Activation Assay