Extended Data Fig. 9: Deletion of p53 protects T cells from STING-mediated proliferation arrest.
(a) Immunoblots of STING in CD4+Cas9+ T cells transduced with sgRNAs targeting STING and p53 at 4 days after retroviral infection. Actin was used as loading control. Representative blots (left) and quantification (right) represented as the mean ± s.e.m. of n = 3 mice and 2 independent experiments. (b) CD80 expression in CD4+Cas9+ T cells transduced with sgRNAs targeting STING and p53, restimulated for 2 days with anti-CD3 + 28 dynabeads and treated or not (vehicle) with 3 μg/ml DMXAA. Representative flow cytometry plots (left) and quantification (right). Data represent the mean ± s.e.m. of at least 3 mice, pooled from 2 independent experiments. (c) CFSE dilution in CD4+Cas9+ T cells transduced with sgRNAs targeting STING and p53, restimulated for 3 days with anti-CD3 + 28 dynabeads and treated or not (vehicle) with STING agonists and idasanutlin (nutlin). Representative flow cytometry plots (left) and quantification (right) of the frequency of proliferating cells. Data represent the mean ± s.e.m. of 3-5 mice, pooled from 2 independent experiments. Statistical analysis by two-tailed, unpaired Student’s t test (a,b) and 2-way ANOVA with Dunnett’s multiple comparisons test (c). Not significant (n.s.) P > 0.05, *P < 0.05, **P < 0.01 and ***P < 0.001. #P < 0.001 between treatment vs vehicle untreated.
