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