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. 2022 Aug 8;7(15):e159430. doi: 10.1172/jci.insight.159430

Figure 5. RNAP3 inhibition suppresses R-loop accumulation and inflammasome activity.

Figure 5

(A) SGM models were treated with ML-60218 (RNAP3 inhibitor, RNAP3i) for 72 hours, and R-loop (S9.6) expression was assessed by flow cytometry (n = 2 each for Tet2 cells and n = 3 each for Srsf2 cells) with representative flow histograms. (B) Representative IF images of R-loop reduction in Tet2–/– immortalized cells treated with RNAP3i (scale bar: 10 μm) and (C) MFI quantitation of immunofluorescence (n = 10 vehicle-treated cells and n = 12 RNAP3i-treated cells). (D) Caspase-1 activity by flow cytometry with representative histograms (n = 3 each). (E) Low-risk MDS BM-MNCs treated in vitro with 10 μM RNAP3i for 72 hours (n = 3), resulting in a decrease in R-loops with representative histogram, ISGs (n = 3 each) (F), and inflammasome markers (caspase-1, IL-1β, and ASC specks (n = 3 each) (G), with representative flow histograms. Data are presented as mean ± SEM. Student’s t test; *P ≤ 0.05, **P ≤ 0.01, ****P ≤ 0.0001.