Extended Data Fig. 3: PARP11 regulates IFNAR1 downregulation in CD8+ T cells.
From: Targeting PARP11 to avert immunosuppression and improve CAR T therapy in solid tumors
a qPCR analysis of mRNA of indicated genes in Jurkat cells treated with adenosine (Ado, 1 mM) for 30 min. Data are shown as mean ± SEM (n = 3 independently treated cell cultures.). Statistical analysis was performed using ordinary one-way ANOVA with Tukey’s multiple comparisons test. ****P < 0.0001. b Flow cytometry analysis of IFNAR1 levels in EL4 cells, in which indicated genes were knocked out by sgRNA mediated CRISPR-Cas9. Data are shown as mean ± SEM. n = 6 independently treated cell cultures. Statistical analysis was performed using ordinary one-way ANOVA with Tukey’s multiple comparisons test. ****P < 0.0001. c Immunoblot and qPCR analysis of efficiency of indicated genes knockout (due to the lack of PARP11-specific antibodies, levels of expression of PARP11 is demonstrated by showing the product of qPCR). Levels of β-actin and β-tubulin are shown as loading controls. d ADP-ribosylation of HA-β-TrCP immunoprecipitated from 293 T cells treated or not with adenosine (Ado, 1 mM, 30 min) as indicated. After treatment, levels of HA-β-TrCP in the whole-cell lysate (WCL) was analyzed by immunoblotting. Normalized amounts of lysate containing comparable levels of HA-β-TrCP were taken into immunoprecipitation with HA antibody, which was then analyzed by immunoblotting using anti-ADP-ribose antibody and HA antibody. It is representative of 3 independent repeats with similar results. e Representative genotyping of Parp11−/− and Ifnar1−/− mice. e Representative flow cytometry data for Fig. 3g, h. f Representative flow cytometry data for Fig. 3j. g Representative flow cytometry data for Fig. 3k.
