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