Fig. 4: ACLY drives CDK13-mediated ccRCC progression and lipid deposition.

a, b Cell viability assessed by CCK-8 assay in 786-O and 769-P cells transfected with CDK13-knockdown (shCDK13), ACLY-overexpressing (oeACLY) or combined vectors versus controls. shCon, scramble shRNA; oeCon, empty vector. a Results in 786-O cells. b Results in 769-P cells. c, d Colony formation assay evaluating proliferation in cells with CDK13/ACLY modulation. c Results in 786-O cells. d Results in 769-P cells. e, f EdU incorporation analysis by flow cytometry in CDK13-silenced and/or ACLY-overexpressing cells. e Results in 786-O cells. f Results in 769-P cells. g, h 786-O and 769-P cells were transfected as in (a, b), and an EdU assay was used to detect cellular proliferation. Scale bar, 50 μm. g Results in 786-O cells. h Results in 769-P cells. i, j ORO staining of lipid accumulation in 786-O and 769-P cells under CDK13/ACLY modulation. Scale bar, 25 μm. i Results in 786-O cells. j Results in 769-P cells. k–n Lipid droplet visualization via BODIPY 493/503 (green) and Nile Red (red) staining in CDK13/ACLY-manipulated cells. k, l BODIPY 493/503 staining (green). k 786-O cells. l 769-P cells. m, n Nile Red staining (red). m 786-O cells. n 769-P cells. Scale bar, 50 μm (applies to all images). o Subcutaneous xenograft tumor formation using 786-O cells with stable CDK13 knockdown (shCDK13), ACLY knockdown (shACLY) or dual knockdown (shCDK13 + shACLY). Representative tumor images (n = 5 per group). p Tumor volume calculated as (length × width²)/2 over 28 days. q Tumor weights post-resection at endpoint. r Nile Red staining of lipid accumulation in xenograft tumors. Scale bar, 50 μm. The data represent mean ± s.e.m. (*P < 0.05, **P < 0.01, ***P < 0.001 versus shCon).