Fig. 2: BATF2 is indispensable for cytoplasmic DNA-mediated IFN-I response in macrophages. | Nature Communications

Fig. 2: BATF2 is indispensable for cytoplasmic DNA-mediated IFN-I response in macrophages.

From: BATF2 is a glutamine-responsive tumour suppressor required for type-I interferon-dependent anti-tumour immunity

Fig. 2

a We confirmed the establishment of Batf2-targeted deletion in mice (left panel). The mRNA levels of Batf2 in BMDM from indicated genotypes are shown (right panel, n = 3 biologically independent samples; data are presented as mean ± SEM; unpaired two-tailed t-test). b BMDM were transfected with 2.0 µg/mL ISD and 1.0 µg/mL poly(I:C) for 8 h, and total mRNA was extracted for bulk RNA sequencing. The transcriptomes were compared between wildtype and Batf2−/− BMDM. Data represent biologically independent samples (n = 2 per group). c GSEA shows the significantly altered pathways between wildtype and Batf2−/− BMDM in response to STING stimulation. d–g BMDM from wildtype and Batf2−/− mice were transfected with 3.0 µg/mL ISD (d), 1.0 µg/mL cGAMP (e), 1.0 µg/mL poly(I:C) (f) or 1.0 µg/mL 5′ppp-dsRNA (g) for 16 h. Total mRNA was extracted for qPCR. The fold changes of Ifnb1, pan-Ifna and Cxcl10 are shown as indicated. Data represent biologically independent experiments (n = 3). h, i Wild-type and Batf2−/− BMDM were transfected with 3.0 µg/mL ISD (h) or 1.0 µg/mL cGAMP (i) for 24 h. The concentration of Ifn-β in the supernatant was measured by ELISA. Data represent biologically independent experiments (n = 3). j Wild-type and Batf2−/− BMDM were incubated with 10.0 µg/mL diABZI for 8 h. Ifn-β in the supernatant was measured by ELISA. Data represent biologically independent experiments (n = 3). k, l Wild-type and Batf2−/− BMDM were transfected with 1.0 µg/mL poly(I:C) (k) or 1.0 µg/mL 5′ppp-dsRNA (l) for 24 h. Ifn-β in the supernatant was quantified by ELISA. Data represent biologically independent experiments (n = 3). m, n Wild-type and Batf2−/− BMDM were transfected with 3.0 µg/mL ISD (m) or 1.0 µg/mL cGAMP (n). Cell lysates were collected at the indicated time points for immunoblot analysis. The experiment was performed in two biologically independent experiments. Comparisons in (d–l) were made using two-way ANOVA followed by multi-comparison tests and data are presented as mean ± SEM. Source data are provided as a Source Data file.

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