Extended Data Fig. 8: Tri-1 suppresses NLRP3 positivity in replicative senescent cells.
a, b, Knockdown of Caspase 1 (a) and GSDMD (b) in senescent IMR90 cells was validated by RT-qPCR. n = 4 biologically independent experiments. c, Expression of IL1β in ER-RAS induced (by 4-OHT) senescent IMR90 cells with or without the knockdown of GSDMD or Caspase 1 was determined by RT-qPCR analysis. n = 3 biologically independent experiments. d, e, Representative images of immunostaining for NLRP3 in ER-RAS induced (by 4-OHT) senescent IMR90 cells with or without the indicated treatments (d). NLRP3 positive cells were quantified (e). n = 3 biologically independent experiments. Scale bars = 10 μm. f, g, Representative images of immunostaining for ASC to visualize inflammasome formation in the indicated control and ER-RAS induced (by 4-OHT) senescent IMR90 cells (f). ASC speck positive cells were quantified (g). n = 3 biologically independent experiments. Nigericin, a known inducer of inflammasome formation, was used as a positive control (10 μM for 4 hours). Scale bars = 10 μm. h, Expression of Txnrd1 in young (4 months) and aged mice (22 months) with or without Tri-1 treatment was determined by RT-qPCR analysis. n = 4 biologically independent mice per group. i, j, Immunoblot of the indicated proteins in the ovary tissues harvested from young (4 months) and aged mice (22 months) (i). The intensity of the indicated proteins was quantified by NIH ImageJ software and normalized against a loading control β-actin expression (j). n = 5 biologically independent mice per group. Data represent mean ± s.e.m. P-values were calculated using a two-tailed t test.
