Fig. 4
LSH is required for p53-mediated lipid metabolism. a A549 cells stably expressing shCon or LSH shRNA were seeded on glass coverslips overnight, and the cells were fixed and stained with red neutral lipid stain. Representative images from three independent experiments are presented (scale bars, 10 μm). b ImageJ was used to analyse the intensity of LipidTox-Red staining. Statistical analysis was performed using Student’s t test. *P < 0.05; **P < 0.01. Error bars represent the SEM of triplicate experiments. c HK1 cells stably expressing Vector, LSH truncations, or LSH were seeded on glass coverslips overnight, and the cells were fixed and stained with red neutral lipid stain. Representative images from three independent experiments are presented (scale bars, 10 μm). d ImageJ was used to analyse the intensity of LipidTox-Red staining. Statistical analysis was performed using Student’s t test. *P < 0.05; **P < 0.01. Error bars represent the SEM of triplicate experiments. e Alterations in lipid metabolism genes were observed in A549 cells stably expressing shCon or LSH shRNA. Cells were harvested and the indicated mRNA levels were determined by real-time PCR. Statistical analysis was performed using Student’s t test. *P < 0.05; **P < 0.01. Error bars represent the SEM of triplicate experiments. f Alterations in lipid metabolism genes were observed in HCT116 p53−/− and HCT116 p53+/+ cells transiently expressing shCon or LSH shRNA. Cells were harvested, and the indicated mRNA levels were determined by real-time PCR. Statistical analysis was performed using Student’s t test. *P < 0.05; **P < 0.01. Error bars represent the SEM of triplicate experiments. g ChIP analysis of p53 target genes in A549 cells transiently depleted of LSH. The enrichment of p53 was assessed using CPT1B, CPT1C, CEL and p21 primers spanning the genomic regions around the TSS. IgG served as an antibody control and p21 as a positive control. Statistical analysis was performed using Student’s t test. *P < 0.05; **P < 0.01. Error bars represent the SEM of triplicate experiments. h Effects of transient overexpression of LSH and p53 in HK1 cells determined by Western blotting for FASN, ACC, and p-ACLY. Representative images from three independent experiments are presented