Figure 2. Inhibition of the mevalonate pathway sensitizes ARID1A-inactivated cells to pyroptosis.

A, ARID1A knockout decreases the cholesterol biosynthesis pathway in OVCA429 cells based on GSEA enrichment analysis of differentially expressed proteins in a previously published proteomic analysis³¹.

B, ARID1A knockout enriches the NLRP3 inflammasome gene signature in OVCA429 cells based on GSEA analysis of differentially expressed genes in a previously published RNA-seq analysis (GEO: GSE131917)⁵⁹.

C, Representative immunofluorescence images of ASC-mCerulean staining in the indicated OVCA429 cells treated with simvastatin (5 μM) or atorvastatin (10 μM). Arrows point to examples of ASC oligo, a marker of inflammasome. Nuclei were visualized by RedDot staining. Scale bar = 20 μm.

D, Quantification of ASC oligomerization positive cells in the indicated treatment groups.

E, Transmission electron microscope images of control and ARID1A knockout OVCA429 cells treated with or without simvastatin (10 μM for 48 hours). The cell surface exhibited disrupted regions and membrane pores (black arrows). Scale bar = 1 μm.

F, Expression of the indicated proteins in control and ARID1A knockout OVCA429 cells treated with or without the indicated concentration of simvastatin was determined by immunoblot. Nigericin treatment was used as a positive control.

G-H, Secretion of IL-1β (G) or IL-18 (H) in control and ARID1A knockout OVCA429 cells treated with or without simvastatin (10 μM for 48 hours) was examined by ELISA. n=3 independent experiments.

I-J, Activity of caspase 1 in control or ARID1A knockout OVCA429 cells treated with or without simvastatin (10 μM) and atorvastatin (10 μM) was examined by flow cytometry (I). And percentages of caspase 1 positive cells were quantified (J). n=3 independent experiments.

K, Expression of the indicated proteins in control and ARID1A knockout OVCA429 cells with or without wildtype ARID1A restoration treated with vehicle control or simvastatin (10 μM for 48 hours) was determined by immunoblot.

L-M, Activity of caspase 1 in control and ARID1A knockout OVCA429 cells with or without wildtype ARID1A restoration treated with vehicle control or simvastatin (10 μM for 48 hours) was examined by flow cytometry (L). And percentages of caspase 1 positive cells were quantified (M). n=3 independent experiments.

N, The prenylation of RhoA and Rac1 in control and ARID1A knockout OVCA429 cells treated with vehicle control, simvastatin (10 μM) with or without supplementation of mevalonate (100 μM) or GGPP (10 μM) for 48 hours were examined by immunoblot.

Error bars represent mean with SEM in D, G, H, J and M. P values were calculated using two-tailed Student t-test in D, G, H, J and M.

See also Figure S2.