eif4g2  (Cell Signaling Technology Inc)

Journal: iScience

Article Title: Translation factor eIF4G2 directs CD8 + T cell lineage commitment by selectively enabling the IL-7 receptor response

doi: 10.1016/j.isci.2026.115313

Figure Lengend Snippet: Eif4g2 expression is upregulated at late stage during T cell development (A) UMAP dimensionality reduction of the first 25 PCs, classifying 12 cell clusters. (B) Dot plot showing marker genes used to identify clusters based on differential gene expression. (C) UMAP plot illustrating the subtypes of the T clusters, color-coded by cell type. (D) Expression ratio of translation initiation factor family members in DP and DN cells. Red bars indicate genes with p < 0.05, black bars indicate genes with p > 0.05. (E) Expression levels of Eif4g2 in DN and DP cells. ∗∗∗ p < 0.001. All data are plotted as mean ± SEM. See also .

Article Snippet: Membranes were blocked with 5% non-fat milk or bovine serum albumin (BSA) in Tris-buffered saline containing 0.1% Tween-20 (TBST) for 1 hour at room temperature and then incubated overnight at 4 °C with the following primary antibodies diluted in blocking buffer: eIF4G2 (CST, 3468S), β-actin (CST, 3700S), eIF4G1 (Proteintech, 15704-1-AP), IL-7Rα (Proteintech, 17626-1-AP), γc (Proteintech, 11409-1-AP), STAT5 (Proteintech, 13179-1-AP), phospho STAT5 (CST, 4322T), STAT6 (Proteintech, 51073-1-AP), mouse phospho STAT6 (CST, 56554S).

Techniques: Expressing, Marker, Gene Expression

Journal: iScience

Article Title: Translation factor eIF4G2 directs CD8 + T cell lineage commitment by selectively enabling the IL-7 receptor response

doi: 10.1016/j.isci.2026.115313

Figure Lengend Snippet: Conditional deletion of Eif4g2 specifically impairs SP thymocyte development (A and B) Validation of eIF4G2 protein deletion by western blot. (A) Total thymocytes. (B) Lysates from sorted thymocyte subsets: double-negative (DN), double-positive (DP), and single-positive (SP) cells. (C) Representative images of thymus from WT and Eif4g2 cKO mice. (D) Total thymocytes numbers from WT and Eif4g2 cKO mice ( n = 3 per group, ns p > 0.05). (E) Flow cytometric analysis of thymocyte populations. (F–H) Relative frequencies and absolute numbers of (F) CD4 SP, (G) CD8 SP, and (H) TCRβ + CD8 + subsets (n = 3–6 mice per group, presented as mean ± SEM. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.001). (I–M) Frequencies of (I) DN, (J) DP cells within total thymocytes, and (K) Foxp3 + cell within CD4 + T cells (n = 5–6 mice per group, presented as mean ± SEM, ns p > 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001); (L and M) Analysis of innate-like T cells in the thymus. (L) Frequencies of NK1.1 + T cells and (M) γδ T cells ( n = 5 mice per group, presented as mean ± SEM. ns p > 0.05, ∗∗∗ p < 0.001). (N–P) Evaluation of peripheral T cells in situ . (N) Absolute numbers of splenic T cells, (O) frequencies of CD44 + CD62L − cells and (P) IFNγ + cells ( n = 3 mice per group, presented as mean ± SEM. ∗ p < 0.05, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001). (Q and R) Cytokine production upon stimulation. Frequencies of (Q) IFNγ + and (R) TNFα + cells among peripheral naive T cells following ex vivo anti-CD3/CD28 stimulation ( n = 3 mice per group, bar graphs show mean ± SEM. ∗ p < 0.05, ∗∗ p < 0.01). Data are representative of at least two independent experiments unpaired Students’ t test was used to perform the statistical analysis. See also .

Article Snippet: Membranes were blocked with 5% non-fat milk or bovine serum albumin (BSA) in Tris-buffered saline containing 0.1% Tween-20 (TBST) for 1 hour at room temperature and then incubated overnight at 4 °C with the following primary antibodies diluted in blocking buffer: eIF4G2 (CST, 3468S), β-actin (CST, 3700S), eIF4G1 (Proteintech, 15704-1-AP), IL-7Rα (Proteintech, 17626-1-AP), γc (Proteintech, 11409-1-AP), STAT5 (Proteintech, 13179-1-AP), phospho STAT5 (CST, 4322T), STAT6 (Proteintech, 51073-1-AP), mouse phospho STAT6 (CST, 56554S).

Techniques: Biomarker Discovery, Western Blot, In Situ, Ex Vivo

Journal: iScience

Article Title: Translation factor eIF4G2 directs CD8 + T cell lineage commitment by selectively enabling the IL-7 receptor response

doi: 10.1016/j.isci.2026.115313

Figure Lengend Snippet: eIF4G2 facilitates CD8 + T lineage commitment after positive selection (A) Flow cytometry gating strategy for WT and Eif4g2 cKO thymocytes based on CD3 and CD69 expression and representative plots for WT and Eif4g2 cKO mice are shown. (B and C) Quantification of the thymocyte subpopulations defined in (A). Bar graphs show the frequencies of each population within total thymocytes ( n = 6, ns p > 0.05, ∗∗∗ p < 0.001, ∗ p < 0.05). (D) Gating strategy to analyze CD4 and CD8 expression within CD3 high CD69 + population. (E) Ratio of CD4 + CD8 − , CD4 + CD8 lo , or CD4 − CD8 + cells in CD3 high CD69 + population ( n = 6, ns p > 0.5, ∗∗∗ p < 0.001). (F) Ratio of CD4 + CD8 − or CD4 − CD8 + cell in CD3 high CD69 - population ( n = 6, ∗ p < 0.5, ∗∗∗ p < 0.001); Data are representative of at least two independent experiments. Bar graphs show mean ± SEM and unpaired Students’ t test was used to perform the statistical analysis.

Article Snippet: Membranes were blocked with 5% non-fat milk or bovine serum albumin (BSA) in Tris-buffered saline containing 0.1% Tween-20 (TBST) for 1 hour at room temperature and then incubated overnight at 4 °C with the following primary antibodies diluted in blocking buffer: eIF4G2 (CST, 3468S), β-actin (CST, 3700S), eIF4G1 (Proteintech, 15704-1-AP), IL-7Rα (Proteintech, 17626-1-AP), γc (Proteintech, 11409-1-AP), STAT5 (Proteintech, 13179-1-AP), phospho STAT5 (CST, 4322T), STAT6 (Proteintech, 51073-1-AP), mouse phospho STAT6 (CST, 56554S).

Techniques: Selection, Flow Cytometry, Expressing

Journal: iScience

Article Title: Translation factor eIF4G2 directs CD8 + T cell lineage commitment by selectively enabling the IL-7 receptor response

doi: 10.1016/j.isci.2026.115313

Figure Lengend Snippet: Eif4g2 deletion specifically ablates the IL-7 response (A and B) Single-cell transcriptomic landscape of thymocytes. UMAP visualization of (A) all thymic cells and (B) the subtypes of T cells, color-coded by cell type. (C) Heatmap showing the expression pattern of 15 core signaling component genes from seven selected KEGG pathways across the cell clusters identified in (A and B). (D–F) Functional response of CD4 + CD8 lo transitional cells to IL-7. Relative mRNA level of (D) Runx3 and (E) Bcl2 following 10 ng/ml IL-7 stimulation ( n = 3, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001). (F) Western blot analysis of key signaling pathway activation with or without IL-7 stimulation. (G) Quantification of cell death in peripheral naive T cells under IL-7 stimulation ex vivo ( n = 3, ∗∗∗∗ p < 0.0001). (H–J) TCR signaling evaluation in DP cells under stimulation with anti-TCRβ/CD2 ex vivo , including (H) representative plots of CD69 expression examination, (I) frequency of CD69 + cells and (J) cell death level ( n = 4, ns p > 0.5). Data are representative of at least two independent experiments. Bar graphs show mean ± SEM and unpaired Students’ t test was used to perform the statistical analysis. See also .

Article Snippet: Membranes were blocked with 5% non-fat milk or bovine serum albumin (BSA) in Tris-buffered saline containing 0.1% Tween-20 (TBST) for 1 hour at room temperature and then incubated overnight at 4 °C with the following primary antibodies diluted in blocking buffer: eIF4G2 (CST, 3468S), β-actin (CST, 3700S), eIF4G1 (Proteintech, 15704-1-AP), IL-7Rα (Proteintech, 17626-1-AP), γc (Proteintech, 11409-1-AP), STAT5 (Proteintech, 13179-1-AP), phospho STAT5 (CST, 4322T), STAT6 (Proteintech, 51073-1-AP), mouse phospho STAT6 (CST, 56554S).

Techniques: Single Cell, Expressing, Functional Assay, Western Blot, Activation Assay, Ex Vivo

Journal: iScience

Article Title: Translation factor eIF4G2 directs CD8 + T cell lineage commitment by selectively enabling the IL-7 receptor response

doi: 10.1016/j.isci.2026.115313

Figure Lengend Snippet: eIF4G2 specifically sustains surface expression of the IL-7 receptor (A–C) Expression of IL-7Rα (CD127). (A) Representative histogram and (B) quantification of CD127 median fluorescence intensity (MFI) on CD4 + CD8 lo transitional cells ( n = 3, ∗∗ p < 0.01). (C) MFI of CD127 on DP, CD4 SP, and CD8 SP thymocytes ( n = 3, ns p > 0.05, ∗∗∗∗ p < 0.0001). (D–F) Expression of the common γc (CD132). (D) Representative histogram and (E) quantification of CD132 MFI on CD4 + CD8 lo transitional cells ( n = 4, ∗ p < 0.05). ppp(F) MFI of CD132 on DP, CD4 SP, and CD8 SP thymocytes ( n = 3, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001). (G and H) Expression of IL-4Rα (CD124) expression detection on CD4 + CD8 lo transitional cells. (G) Representative histogram and (H) quantification of CD124 MFI on CD4 + CD8 lo transitional cells ( n = 3, ns p > 0.05). (I and J) Expression of GP130 on CD4 + CD8 lo transitional cells. (I) Representative histogram and (J) quantification of GP130 MFI on CD4 + CD8 lo transitional cells ( n = 3, ∗∗ p < 0.01). (K) Quantification of CD124 MFI on DP, CD4 SP and CD8 SP ( n = 3, ns p > 0.05). (L–N) Expression of IL-2Rα (CD25). (L) Representative histogram and (M) quantification of CD25 MFI on CD4 + CD8 lo transitional cells ( n = 3, ns p > 0.05). (N) MFI of CD25 on DP, CD4 SP, and CD8 SP subsets ( n = 3, ns p > 0.05). Data are representative of at least two independent experiments. Bar graphs show mean ± SEM and unpaired Students’ t test was used to perform the statistical analysis.

Article Snippet: Membranes were blocked with 5% non-fat milk or bovine serum albumin (BSA) in Tris-buffered saline containing 0.1% Tween-20 (TBST) for 1 hour at room temperature and then incubated overnight at 4 °C with the following primary antibodies diluted in blocking buffer: eIF4G2 (CST, 3468S), β-actin (CST, 3700S), eIF4G1 (Proteintech, 15704-1-AP), IL-7Rα (Proteintech, 17626-1-AP), γc (Proteintech, 11409-1-AP), STAT5 (Proteintech, 13179-1-AP), phospho STAT5 (CST, 4322T), STAT6 (Proteintech, 51073-1-AP), mouse phospho STAT6 (CST, 56554S).

Techniques: Expressing, Fluorescence

Journal: iScience

Article Title: Translation factor eIF4G2 directs CD8 + T cell lineage commitment by selectively enabling the IL-7 receptor response

doi: 10.1016/j.isci.2026.115313

Figure Lengend Snippet: eIF4G2 post-transcriptionally sustains γc expression via its mRNA UTRs (A–C) Analysis in primary CD4 + CD8 lo transitional thymocytes. (A) Western blot analysis of γc and IL-7Rα protein levels. (B and C) Quantitative RT-PCR analysis of Il2rg (B) and Il7r (C) mRNA levels ( n = 3, ns p > 0.05, ∗ p < 0.05). (D–H) Mechanistic dissection in 293T cells. (D) Western blot of γc protein in control and EIF4G2 knockdown 293T cells transfected with an IL2RG coding sequence construct containing its native 5′ and 3′ UTRs. (E) Corresponding IL2RG mRNA levels measured by RT-qPCR ( n = 3, ns p > 0.05) . (F and G) Assessment of γc protein stability ( n = 3, ns p > 0.05). (F) Representative western blots and (G) quantification of γc protein levels over time following cycloheximide (CHX) treatment in si-control and si- EIF4G2 293T cells ( n = 3, ns p > 0.05). (H) Western blot of γc protein in si-control and si- EIF4G2 293T cells transfected with an IL2RG CDS construct lacking UTRs. Data are representative of at least two independent experiments. Bar graphs show mean ± SEM and unpaired Students’ t test was used to perform the statistical analysis.

Article Snippet: Membranes were blocked with 5% non-fat milk or bovine serum albumin (BSA) in Tris-buffered saline containing 0.1% Tween-20 (TBST) for 1 hour at room temperature and then incubated overnight at 4 °C with the following primary antibodies diluted in blocking buffer: eIF4G2 (CST, 3468S), β-actin (CST, 3700S), eIF4G1 (Proteintech, 15704-1-AP), IL-7Rα (Proteintech, 17626-1-AP), γc (Proteintech, 11409-1-AP), STAT5 (Proteintech, 13179-1-AP), phospho STAT5 (CST, 4322T), STAT6 (Proteintech, 51073-1-AP), mouse phospho STAT6 (CST, 56554S).

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Dissection, Control, Knockdown, Transfection, Sequencing, Construct

Journal: Journal of Advanced Research

Article Title: Endothelial cells secrete small extracellular vesicles to promote neuron endoplasmic reticulum stress injury via miR-146a-5p/Eif4g2 axis in ischemic stroke

doi: 10.1016/j.jare.2025.07.009

Figure Lengend Snippet: Eif4g2 was a potential target of miR-146a-5p and necessary for neuron survival. (A) Venn diagram illustrating the potential targets of miR-146a-5p identified through TargetScan, miRDB, and starBase databases. Online queries revealed 12 overlapping potential binding partners of miR-146a-5p. (B, C) Expression levels of 12 potential binding partners of miR-146a-5p following treatment with OGD/R ECs-sEVs and miR-146a-5p mimic, n = 3. (D) Predicted putative seed-matching sites between miR-146a-5p and Eif4g2, along with luciferase reporter assay results, n = 5. (E) Representative images of TUNEL staining, n = 5. (F-H) Representative western blot analysis images and statistical results of Bcl-2/Bax and cleaved caspase3/caspase3 in HT22 cells and mice brain with Eif4g2 siRNA, n = 3. (I, J) Representative images of cerebral blood flow and Nissl staining of the hippocampus after Eif4g2 downregulation, n = 5. Data are presented as the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001 vs . NC group. ns: no significance.

Article Snippet: This blocking step was succeeded by an overnight incubation at 4 °C with a mix of primary antibodies, namely: Bcl-2 (Abclonal), Bax (Proteintech), cleaved caspase-3 (Abcam), caspase-3 (Abclonal), β-actin (Abclonal), Eif4g2 (Proteintech), ATF6 (Proteintech), GRP78 (Proteintech), and CHOP (Abcam).

Techniques: Binding Assay, Expressing, Luciferase, Reporter Assay, TUNEL Assay, Staining, Western Blot

Journal: Journal of Advanced Research

Article Title: Endothelial cells secrete small extracellular vesicles to promote neuron endoplasmic reticulum stress injury via miR-146a-5p/Eif4g2 axis in ischemic stroke

doi: 10.1016/j.jare.2025.07.009

Figure Lengend Snippet: MiR-146a-5p regulated GRP78/ATF6/CHOP pathway by targeting Eif4g2. (A, B) Western blot analysis images of Eif4g2, GRP78, ATF6, and CHOP in brain of mice after miR-146a-5p mimic and inhibitor. (C, D) Western blot analysis images of Eif4g2, GRP78, ATF6, and CHOP in mice brain after miR-146a-5p agomir and antagomir. Data are presented as the mean ± SD, n = 3. * P < 0.05, ** P < 0.01 vs . NC group. & P < 0.05, && P < 0.01 vs .

Article Snippet: This blocking step was succeeded by an overnight incubation at 4 °C with a mix of primary antibodies, namely: Bcl-2 (Abclonal), Bax (Proteintech), cleaved caspase-3 (Abcam), caspase-3 (Abclonal), β-actin (Abclonal), Eif4g2 (Proteintech), ATF6 (Proteintech), GRP78 (Proteintech), and CHOP (Abcam).

Techniques: Western Blot

Journal: Journal of Advanced Research

Article Title: Endothelial cells secrete small extracellular vesicles to promote neuron endoplasmic reticulum stress injury via miR-146a-5p/Eif4g2 axis in ischemic stroke

doi: 10.1016/j.jare.2025.07.009

Figure Lengend Snippet: Eif4g2 regulated neuron injury via endoplasmic reticulum stress. (A) Heatmap of RNA-seq in HT22 with Eif4g2 siRNA. Red represents up-regulated genes and green represents downregulated genes, n = 5. (B) Bubble plot of enriched GO terms. (C) Representative endoplasmic reticulum morphological changes in HT22 cells and brain after Eif4g2 siRNA. (D-E) Representative western blot images showing relative protein levels of GRP78, ATF6, and CHOP in HT22 cells and brain after Eif4g2 siRNA, n = 3. Data are presented as the mean ± SD. * P < 0.05 vs . NC-siRNA group.

Article Snippet: This blocking step was succeeded by an overnight incubation at 4 °C with a mix of primary antibodies, namely: Bcl-2 (Abclonal), Bax (Proteintech), cleaved caspase-3 (Abcam), caspase-3 (Abclonal), β-actin (Abclonal), Eif4g2 (Proteintech), ATF6 (Proteintech), GRP78 (Proteintech), and CHOP (Abcam).

Techniques: RNA Sequencing, Western Blot

Journal: Journal of Advanced Research

Article Title: Endothelial cells secrete small extracellular vesicles to promote neuron endoplasmic reticulum stress injury via miR-146a-5p/Eif4g2 axis in ischemic stroke

doi: 10.1016/j.jare.2025.07.009

Figure Lengend Snippet: NBP protected neuron injury via targeting miR-146a-5p in MCAO/R rats. (A) Three-dimensional structural representation of miR-146a-5p binding sites for molecular docking analysis. (B) Schematic representation of the NBP binding partner of miR-146a-5p available for prediction. (C, D) MST and SPR detection for the binding affinity of NBP to pre- miR-146. (E-G) RT-qPCR analysis of miR-146a-5p expression in OGD/R ECs, sEVs, and brain after NBP treatment, n = 8. (H) Representative western blot analysis images of Eif4g2, GRP78, ATF6, and CHOP in MCAO/R rats after NBP treatment, n = 3. Data are presented as the mean ± SD. & P < 0.05, && P < 0.01, &&& P < 0.001 vs . OGD/R ECs or MCAO/R group.

Article Snippet: This blocking step was succeeded by an overnight incubation at 4 °C with a mix of primary antibodies, namely: Bcl-2 (Abclonal), Bax (Proteintech), cleaved caspase-3 (Abcam), caspase-3 (Abclonal), β-actin (Abclonal), Eif4g2 (Proteintech), ATF6 (Proteintech), GRP78 (Proteintech), and CHOP (Abcam).

Techniques: Binding Assay, Quantitative RT-PCR, Expressing, Western Blot

Journal: bioRxiv

Article Title: eIF4G2-dependent translation restrains pancreatic cancer progression

doi: 10.64898/2026.03.14.711799

Figure Lengend Snippet: (A) Schematic of the in vivo CRISPR-Cas9 screening workflow, from library transduction of KPC PDAC cells to tumor harvest in 5 cages of 10-week-old mice. (B) Correlation of gene essentiality scores from the in vitro portion of the screen (beta scores from MAGeCK-MLE, Day 17 vs. Day 0) with pan-cancer essentiality scores from the DepMap database. (C) Reduced correlation of in vivo gene essentiality scores (beta scores, endpoint tumors vs. Day 17) with DepMap scores. (D) Changes in gRNA enrichment between the in vitro outgrowth (Day 17) and in vivo tumors, known tumor suppressors (e.g., Cdkn2b, Pten ) and oncogenes (e.g., Kras, Myc ) are highlighted. (E) Normalized read counts for four independent gRNAs targeting Eif4g2 , showing their strong enrichment in endpoint tumors relative to the Day 17 engraftment-ready cell population. Data are represented as the median of all primary tumors.

Article Snippet: The following primary antibodies were used at 1:1000 dilution: eIF4G2 (D88B6) [Cell Signaling #5169, AB_10622189], eIF4A (C32B4)[Cell Signaling# 2013, AB_2097363], 4EBP1[Cell Signaling# 9452, AB_331692], pS6(D68F8)[Cell Signaling # 5364, AB_10694233], S6 (54D2) [Cell Signaling # 2317, AB_2238583], eIF4E(C46H6) [Cell Signaling # 2067, AB_2097675], PTEN (138G6) [Cell Signaling # 9559, AB_390810], CREBBP(D6C5)[Cell Signaling # 7389, AB_2616020].

Techniques: In Vivo, CRISPR, Transduction, In Vitro

Journal: bioRxiv

Article Title: eIF4G2-dependent translation restrains pancreatic cancer progression

doi: 10.64898/2026.03.14.711799

Figure Lengend Snippet: (A) Distribution of log-transformed sgRNA read counts. Other metrics for screen quality are also shown such as read mapping rates (average > 85%), sgRNA dropout, and uniform read distribution (average Gini index ≈ 0.1). (B) Distribution of sgRNA log2-fold change for the in vitro (Day 17 vs. Day 0) and in vivo (endpoint tumors vs. Day 17) portions of the screen. (C) Correlation matrix of sgRNA read counts across different screening conditions. (D) The correlation of the sgRNA fold change from our in vivo screen with the gene score generated from the PDA in vivo screen performed in the Birsoy lab . (E) The fold change of sgRNAs and volcano plot of the in vitro screen results (Day 17 vs. Day 0), the enrichment of Eif4g2 is highlighted. (F) Normalized read counts for four independent sgRNAs targeting Eif4g2 , between the Day 0 baseline and the Day 17 cell population.

Article Snippet: The following primary antibodies were used at 1:1000 dilution: eIF4G2 (D88B6) [Cell Signaling #5169, AB_10622189], eIF4A (C32B4)[Cell Signaling# 2013, AB_2097363], 4EBP1[Cell Signaling# 9452, AB_331692], pS6(D68F8)[Cell Signaling # 5364, AB_10694233], S6 (54D2) [Cell Signaling # 2317, AB_2238583], eIF4E(C46H6) [Cell Signaling # 2067, AB_2097675], PTEN (138G6) [Cell Signaling # 9559, AB_390810], CREBBP(D6C5)[Cell Signaling # 7389, AB_2616020].

Techniques: Transformation Assay, In Vitro, In Vivo, Generated

Journal: bioRxiv

Article Title: eIF4G2-dependent translation restrains pancreatic cancer progression

doi: 10.64898/2026.03.14.711799

Figure Lengend Snippet: (A) Immunoblot analysis of eIF4G2 expression in bulk tumor lysates. (B) Mass of pancreatic tumors at the 5-week endpoint after orthotopic implantation of control (sg Rosa ) and eIF4G2-deficient (sg Eif4g2 ) KPC cells. sg Eif4g2-2 and sg Eif4g2-4 are two sgRNAs targeting Eif4g2 . (C) Immunohistochemical staining and quantification of phospho-histone H3 in orthotopic PDA tumors. (D-F) Representative H&E images (D) and pathological assessments of sg Rosa and sg Eif4G2 tumor sections based on differentiation status (E) and glandular morphology (F). (G) Number of metastatic lesions across different organ sites. Error bars in this Fig. are means ± SDs. Student’s t-test was performed. ns (not significant) for P ≥ 0.05, * (one asterisk) for P < 0.05, ** (two asterisks) for P < 0.01, *** (three asterisks) for P < 0.001, and **** (four asterisks) for P < 0.0001.

Article Snippet: The following primary antibodies were used at 1:1000 dilution: eIF4G2 (D88B6) [Cell Signaling #5169, AB_10622189], eIF4A (C32B4)[Cell Signaling# 2013, AB_2097363], 4EBP1[Cell Signaling# 9452, AB_331692], pS6(D68F8)[Cell Signaling # 5364, AB_10694233], S6 (54D2) [Cell Signaling # 2317, AB_2238583], eIF4E(C46H6) [Cell Signaling # 2067, AB_2097675], PTEN (138G6) [Cell Signaling # 9559, AB_390810], CREBBP(D6C5)[Cell Signaling # 7389, AB_2616020].

Techniques: Western Blot, Expressing, Control, Immunohistochemical staining, Staining

Journal: bioRxiv

Article Title: eIF4G2-dependent translation restrains pancreatic cancer progression

doi: 10.64898/2026.03.14.711799

Figure Lengend Snippet: (A, B) Proliferation of control (sg Rosa ) and eIF4G2-deficient (sg Eif4g2 ) KPC cells measured by CellTiter-Glo (A) and EdU incorporation (B). (C) Viability of control and eIF4G2-deficient KPC cells upon increasing concentrations of eIF4A inhibitor (CR-1-31-B) for 72 hours. (D) L-Azidohomoalanine (AHA) incorporation in sg Rosa and sg Eif4g2 KPC cells was analyzed by immunoblot, with cells treated with cycloheximide serving as negative controls. (E) OP-Puro incorporation was similarly analyzed by immunoblot, with cycloheximide-treated cells as controls. (F)Polysome profiles of control and eIF4G2-deficient KPC cells. Absorbance at 254 nm is plotted as a function of sedimentation. (G and H) Representative immunoblot analysis of the abundance of eIF4F components (I) and downstream substrates of mTOR activation (J). Error bars in this Fig. are means ± SDs. Student’s t-test was performed. ns (not significant) for P ≥ 0.05, * (one asterisk) for P < 0.05, ** (two asterisks) for P < 0.01, *** (three asterisks) for P < 0.001, and **** (four asterisks) for P < 0.0001.

Article Snippet: The following primary antibodies were used at 1:1000 dilution: eIF4G2 (D88B6) [Cell Signaling #5169, AB_10622189], eIF4A (C32B4)[Cell Signaling# 2013, AB_2097363], 4EBP1[Cell Signaling# 9452, AB_331692], pS6(D68F8)[Cell Signaling # 5364, AB_10694233], S6 (54D2) [Cell Signaling # 2317, AB_2238583], eIF4E(C46H6) [Cell Signaling # 2067, AB_2097675], PTEN (138G6) [Cell Signaling # 9559, AB_390810], CREBBP(D6C5)[Cell Signaling # 7389, AB_2616020].

Techniques: Control, Western Blot, Sedimentation, Activation Assay

Journal: bioRxiv

Article Title: eIF4G2-dependent translation restrains pancreatic cancer progression

doi: 10.64898/2026.03.14.711799

Figure Lengend Snippet: (A) Pairs of ribosome-protected footprint (RPF) and RNA-Seq libraries were prepared from each sample. The plot shows the distribution of RPM (reads per million) normalized gene counts in each of the libraries. (B) This plot shows the location of scaled read ends that have been corrected for their estimated P-site (the second binding site for tRNA in the ribosome) from the ribosome-protected footprint (RPF) libraries. This correction is to show where ribosomes are directly binding, as the sequenced read ends have an overhang past the ribosome complex. RPF libraries are expected to show a trinucleotide periodicity. (C) RPF reads will accumulate where a ribosome pauses or stalls during translation. This plot shows putative stall locations for each transcript in the transcriptome, where stalls were detected through the use of a Kolmogorov-Smirnov (KS) test on normalized cumulative P-site densities. This analysis is performed per transcript, so the listed distances refer to the distance in transcriptomic space, not genomic space. (D) This scatter plot shows the mean codon usage in each condition, where codon usage is defined as the scaled normalized frequency that P-sites were found to be enriched in each codon. Each dot is a different codon, colored by whether it is a start, stop, or an amino acid-coding codon. Off-diagonal codons can indicate differential codon usage between the two conditions. Codon usage was determined using riboWaltz. If a gene has multiple transcripts, only the longest transcript is included in this analysis. (E) qRT-PCR analysis of Pten mRNA distribution across polysome fractions. (F) immunoblot analysis of PTEN expression in control (sg Rosa ) and eIF4G2-deficient (sg Eif4g2 ) bulk tumor lysates. (G) Immunoblot analysis of PTEN expression in sg Rosa and sg Pten KPC cells. (H) Mass of pancreatic tumors from in sg Rosa and sg Pten KPC cells 5 weeks after orthotopic implantation. (I) Immunoblot analysis of eIF4G2 expression in control (sg Rosa ) and Pten -deficient (sg Pten ) bulk tumor lysates. (J) qRT-PCR analysis of Crebbp mRNA distribution across polysome fractions. (K) Volcano plot of eIF4G2-dependent transcriptional changes. (L) KRT14 and PTEN expression in sg Rosa and sg Pten bulk tumor lysates. PTEN and HSP90 immunoblots are also used in panel (I). Error bars in this Fig. are means ± SDs. Student’s t-test was performed. ns (not significant) for P ≥ 0.05, * (one asterisk) for P < 0.05, ** (two asterisks) for P < 0.01, *** (three asterisks) for P < 0.001, and **** (four asterisks) for P < 0.0001.

Article Snippet: The following primary antibodies were used at 1:1000 dilution: eIF4G2 (D88B6) [Cell Signaling #5169, AB_10622189], eIF4A (C32B4)[Cell Signaling# 2013, AB_2097363], 4EBP1[Cell Signaling# 9452, AB_331692], pS6(D68F8)[Cell Signaling # 5364, AB_10694233], S6 (54D2) [Cell Signaling # 2317, AB_2238583], eIF4E(C46H6) [Cell Signaling # 2067, AB_2097675], PTEN (138G6) [Cell Signaling # 9559, AB_390810], CREBBP(D6C5)[Cell Signaling # 7389, AB_2616020].

Techniques: RNA Sequencing, Binding Assay, Quantitative RT-PCR, Western Blot, Expressing, Control

Journal: bioRxiv

Article Title: eIF4G2-dependent translation restrains pancreatic cancer progression

doi: 10.64898/2026.03.14.711799

Figure Lengend Snippet: (A) Volcano plot showing changes in translation efficiency in sg Eif4g2 vs sg Rosa KPC cells. (B) Immunoblot analysis of PTEN expression in control (sg Rosa ) and eIF4G2-deficient (sg Eif4g2 ) KPC cells. (C) Immunoblot analysis of PTEN expression in two independent control (sg Rosa ) and eIF4G2-deficient (sg Eif4g2 ) patient-derived PDA cell lines. (D) Representative H&E images from sg Rosa and sg Pten tumor sections. (E) Pathological assessment of sg Rosa and sg Pten tumor sections based on glandular features and differentiation status. (F) Representative immunoblots of CREBBP in control (sg Rosa ) and eIF4G2-deficient (sg Eif4g2 ) KPC cells. (G) Gene set enrichment analysis of transcripts upregulated upon eIF4G2 loss. (H) Representative immunoblot analysis of KRT14 expression in control (sg Rosa ) and eIF4G2-deficient (sg Eif4g2 ) bulk tumor lysates. Error bars in this Fig. are means ± SDs. Student’s t-test was performed. ns (not significant) for P ≥ 0.05, * (one asterisk) for P < 0.05, ** (two asterisks) for P < 0.01, *** (three asterisks) for P < 0.001, and **** (four asterisks) for P < 0.0001.

Article Snippet: The following primary antibodies were used at 1:1000 dilution: eIF4G2 (D88B6) [Cell Signaling #5169, AB_10622189], eIF4A (C32B4)[Cell Signaling# 2013, AB_2097363], 4EBP1[Cell Signaling# 9452, AB_331692], pS6(D68F8)[Cell Signaling # 5364, AB_10694233], S6 (54D2) [Cell Signaling # 2317, AB_2238583], eIF4E(C46H6) [Cell Signaling # 2067, AB_2097675], PTEN (138G6) [Cell Signaling # 9559, AB_390810], CREBBP(D6C5)[Cell Signaling # 7389, AB_2616020].

Techniques: Western Blot, Expressing, Control, Derivative Assay

Journal: bioRxiv

Article Title: eIF4G2-dependent translation restrains pancreatic cancer progression

doi: 10.64898/2026.03.14.711799

Figure Lengend Snippet: (A) Boxplots showing Z-score–based expression signature scores for Basal and Classical PDAC programs in control (sg Rosa ) and EIF4G2 knockout (sg Eif4g2 ) cells. (B) Boxplots of Epithelial, Mesenchymal, and EMT index scores comparing control and eIF4G2 knockout cells. (C) EIF4G2 protein activity analysis in primary versus metastatic tumors from the UNC PDAC cohort . Differences between groups were evaluated using an unpaired, two-sided Wilcoxon rank-sum test; p -value indicated. (D) Kaplan-Meier analysis of overall survival of patients stratified by EIF4G2 activity (High > 0, Low ≤ 0) in the UNC PDAC cohort . Survival probability was estimated nonparametrically and compared with a two-sided log-rank test; the number at risk is shown at the bottom.

Article Snippet: The following primary antibodies were used at 1:1000 dilution: eIF4G2 (D88B6) [Cell Signaling #5169, AB_10622189], eIF4A (C32B4)[Cell Signaling# 2013, AB_2097363], 4EBP1[Cell Signaling# 9452, AB_331692], pS6(D68F8)[Cell Signaling # 5364, AB_10694233], S6 (54D2) [Cell Signaling # 2317, AB_2238583], eIF4E(C46H6) [Cell Signaling # 2067, AB_2097675], PTEN (138G6) [Cell Signaling # 9559, AB_390810], CREBBP(D6C5)[Cell Signaling # 7389, AB_2616020].

Techniques: Expressing, Control, Knock-Out, Activity Assay

Journal: bioRxiv

Article Title: eIF4G2-dependent translation restrains pancreatic cancer progression

doi: 10.64898/2026.03.14.711799

Figure Lengend Snippet: (A) EIF4G2 mRNA abundance based on a recently published morpho-biotype framework for PDA, relative to well-differentiated lesions.

Article Snippet: The following primary antibodies were used at 1:1000 dilution: eIF4G2 (D88B6) [Cell Signaling #5169, AB_10622189], eIF4A (C32B4)[Cell Signaling# 2013, AB_2097363], 4EBP1[Cell Signaling# 9452, AB_331692], pS6(D68F8)[Cell Signaling # 5364, AB_10694233], S6 (54D2) [Cell Signaling # 2317, AB_2238583], eIF4E(C46H6) [Cell Signaling # 2067, AB_2097675], PTEN (138G6) [Cell Signaling # 9559, AB_390810], CREBBP(D6C5)[Cell Signaling # 7389, AB_2616020].

Techniques: