Fig 1: Dispersing PKM2 aggregates impedes cellular senescence.a Model of screening for compounds dispersing PKM2 aggregates. b Immunofluorescent imaging of PKM2 aggregates in HeLa cells stably expressed sfcherry-PKM2 and treated with 2 μM etoposide (ETO) together with DMSO/K35/K27 for two days. Scale bar, 5 μm. c, d Representative images (c) and quantification (d) of SA-β-gal staining in HeLa or HEK 293T cells treated with 10 μM etoposide for 24 h and released for three days together with DMSO or K35 (50 μM). Scale bar, 100 μm. n = 5 (randomly captured images), one-way ANOVA. e Immunoblotting in HEK 293T cells treated with 2 μM ETO for three days together with concentration gradient of K35. f qPCR analysis in HEK 293T cells treated with 10 μM ETO for 24 h and released for indicated days together with DMSO or K35 (50 μM). n = 3 (biological replicates), Two-tailed unpaired t-test, P = 0.0012 (p21: Day 2), P = 0.0006 (p21: Day 3); P = 0.0055 (IL1A: Day 2), P = 0.0020 (IL1A: Day 3); P = 0.0004 (IL6: Day 2), P = 0.0001 (IL6: Day 3); P = 0.0295 (IL1B: Day 2), P = 0.0169 (IL1B: Day 3). g Immunoblotting in HEK 293T cells treated with 10 μM ETO for 24 h and released for three days together with DMSO, K35 or K35 analogs. h, i Representative images (h) and quantification (i) of SA-β-gal staining in fibroblasts 2BS (P25) exposed to 20 μM ETO for 24 h and released for three days together with DMSO, 50 μM K35/K27. Scale bar, 100 μm. n = 6 (randomly captured images), one-way ANOVA. j Immunoblotting in fibroblasts 2BS constantly treated with DMSO, 25 μM K35/K27. k, l Representative images (k) and quantification (l) of SA-β-gal staining in fibroblasts 2BS cultured with DMSO, 25 μM K35/K27 constantly. Scale bar, 100 μm. n = 4 (randomly captured images), one-way ANOVA. m, n Representative confocal images (m) and quantification (n) of EdU staining proliferation assay in HeLa cells treated with 10 μM ETO for 24 h and released for three days together with DMSO, 50 μM K35/K27. Scale bar, 50 μm. n = 5 (randomly captured images), one-way ANOVA. o Quantification of lactate in the culture medium of HeLa cells exposed to 10 μM ETO for 24 h and released for three days together with DMSO, DMSO, 50 μM K35/K27. n = 3 (biological replicates), one-way ANOVA. p Quantification of lactate in the culture medium of young or senescent fibroblasts 2BS exposed to DMSO, 25 μM K35/K27. n = 3 (biological replicates), one-way ANOVA, P = 0.0016 (Young: DMSO vs. K35), P = 0.0001 (Young: DMSO vs. K27); P = 0.0050 (Senescent: DMSO vs. K35). q Measurement of PKM2 enzymatic activity via LDH-coupled kinetic assay in young or senescent fibroblasts 2BS exposed to DMSO, 25 μM K35/K27. n = 3 (technical triplicates), one-way ANOVA, P = 0.0002 (Senescent vs. Senescent + K35). r Measurement of PKM2 enzymatic activity with a commercial kit (K709-100) in young or senescent fibroblasts 2BS exposed to DMSO, 25 μM K35/K27. n = 3 (biological replicates), one-way ANOVA, P = 0.0090 (Young: DMSO vs. K35), P = 0.0007 (Young: DMSO vs. K27); P = 0.0002 (Senescent: DMSO vs. K35). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Error bars represent SEM. Source data are provided as a Source Data file.