Skip to main content
. 2021 Dec 28;23(1):311. doi: 10.3390/ijms23010311

Figure 4.

Figure 4

Evidence of p53-independent Fas/FasL upregulation and induction of apoptosis in E2f2−/− T lymphocytes: (A) Apoptotic death in TCR-stimulated T cells derived from lymph nodes of WT, E2f2−/−, p53−/− and E2f2−/−/p53−/− mice, assessed by Annexin V-FITC and PI staining followed by FACS analysis. Results are expressed as the percentage of cells undergoing early apoptosis (mean ± SD) from 3 independent experiments. (B) Representative Western blot analysis of cleaved Caspase 3, Parp-1, Fas, and FasL in extracts prepared from freshly purified WT, E2f2−/− p53−/− and E2f2/p53−/− (DKO) T cells unstimulated or after stimulation with anti-CD3 during the indicated time. Expression of β-actin was used as loading control. Numbers below the bands correspond to the relative densitometric values, expressed as fold-over WT at 24 h. Similar results were obtained in 3 experiments. (C) Reverse transcription qPCR analysis of Bax, Fas, and FasL in WT, E2f2−/−, p53−/−, and E2f2/p53−/− purified T cells unstimulated or after stimulation with anti-CD3 during the indicated time. Eef1a1 was used as normalization control. Results are expressed as fold-over WT at 0 h (mean ± SD) from 5 independent experiments. ** p < 0.005, * p < 0.05, ns = non-significant.