Fig. 2: Srf-null memory cells cannot mount an effective recall response. | Nature Communications

Fig. 2: Srf-null memory cells cannot mount an effective recall response.

From: IL-2 delivery to CD8+ T cells during infection requires MRTF/SRF-dependent gene expression and cytoskeletal dynamics

Fig. 2

A Protocol for post-infection inactivation of Srf. For post-infection, WT OT-I TamCre (CD45.1) and OT-I Srf  f/f TamCre (CD45.2) CD8+ T cells were co-injected at a ratio 1:1 into CD45.1/CD45.2 hosts, which were infected with rLM-OVA the next day. OT-I Srf−/−(post): four days after infection, animals were fed tamoxifen to induce Srf inactivation. B Immunoblotting analysis of Srf following tamoxifen administration. C Expansion of WT OT-I TamCre and OT-I Srf−/−(post) CD8+ T cells in blood. Data show mean values ± SEM, 3 mice per time point. Representative of two independent experiments. Right, percentages of OT-I WT and OT-I Srf−/− CD8+ cells at day 8 after infection in the blood of animals. OT-I Srf−/−(post): four days after infection, animals were fed tamoxifen to induce Srf inactivation. OT-I Srf−/−(pre): animals transferred with Srf-null cells as in Fig. 1A. Data from 3 independent experiments, one animal per data point and paired two-tailed t test. D A schematic of OT-I WT (CD45.1) and OT-I Srf−/−(post) (CD45.2) memory cell populations purified at day 50 after primary LM-OVA infection from spleen of animals treated with the post-infection deletion regime as in Fig. 2A and transferred at 1:1 ratio to a secondary CD45.1/CD45.2 host, which was subjected to infection the following day. E Expansion profiles, percentages and absolute cell counts of OT-I WT and OT-I Srf−/−(post) memory cells are compared 6 days after infection. Left, representative plot of input and expanded CD8 + T cells. Right, data from 4 individual mice after infection at day 6. Dot plot represent individual mice, Mean values ± SEM are shown; Statistical significance: paired two-tailed t test. F IL-2Rα cell surface expression in OT-I WT (blue) and OT-I Srf−/−(post) (red) cells 6 days after memory immune response, performed as in (D).Top, representative plot of IL-2Rα expression. Bottom, data from 4 individual mice after infection at day 6. Datapoints with Mean values ± SEM are shown. Statistical significance: paired t test. Source data are provided as a Source Data file.

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