Extended Data Fig. 3: SK56 inhibits pyroptosis in vitro.

a, The atom frequency distribution histogram of the number at the interface (with H) involved in nearly 40,000 pairs of interaction interfaces. About 90% of the interfaces involved in the PDB database contain fewer than 350 atoms. b, The variation in loss during model training (total of 153k steps). c, SDS-PAGE image showing the prokaryotic expression and purification of SK56 (including tag cleavage). d, Live-cell imaging showing pyroptosis-induced cell rupture delay in BMDM cells treated with or without SK56 (15 µM). n = 3 repeats. Scale bar, 50 µm. e, ATP assay showing concentration-dependent suppression of pyroptosis by SK56 in BMDM cells. n = 3 repeats. f, 3D imaging showing THP-1 cell pyroptosis treated with or without SK56. Scale bar, 20 µm. g-h, ATP assay showing IC₅₀ of SK56 in classical pyroptosis pathway (g) in THP-1 cells or nonclassical pyroptosis pathway in BMDM cells (h) at 3 h post-induction. n = 3 repeats. i, ELISA results showing the release of IL-1β during nonclassical pyroptosis treated with or without SK56 in BMDM cells. n = 3 repeats. j-m, Cell death assays showing HT29 (j, necrosis, TSZ: TNF-α + SM-164 + Z-VAD-FMK, n = 3 repeats), RAW264.7 (k, apoptosis, n = 4 repeats), AC16 (l, cuproptosis, n = 3 repeats), and MDA-MB-231 (m, ferroptosis, n = 3 repeats) cell responses to SK56. All graphs present mean ± s.d., and P values were determined using two-tailed Student’s t-test, NS (P > 0.05, not significant).