Fig. 4

DLAT enhances FASN expression by upregulating SREBP1. (A-B) The effects of DLAT knockdown on SREBP1, SREBP2, and chREBP mRNA and protein expression levels were examined in OC cells by qRT-PCR and western blot analysis, respectively. SKOV3 or OVCAR3 cells were transduced with sh-NC, sh-DLAT#1, or sh-DLAT#2 lentiviruses. Cells not transduced with lentivirus were used as controls. GAPDH was selected as the internal control. (C) Scatter plot analysis showing the correlation between DLAT and SREBP1 mRNA expression levels in 70 OC tissues. (D-G) SKOV3 or OVCAR3 cells were divided into the following treatment groups: sh-NC, sh-DLAT, sh-DLAT + EV (empty vector) and sh-DLAT + oeSREBP1. (D) Western blot analysis of SREBP1, ACLY, FASN, ACC, and SCD protein expression levels in the indicated treatment groups. GAPDH was used as a loading control. (E-F) Cellular contents of triglyceride (E) and cholesterol (F) were measured in the indicated cells. (G) Nile Red staining was used to measure cellular neutral lipids in the indicated cells. Nuclei were stained with Hoescht (blue). Scale bar = 50 μm. (H) qRT-PCR analysis of LDHA and LDHB mRNA expression levels in SKOV3 and OVCAR3 cells with indicated treatments. Data are presented as the mean ± SD, n = 3. For statistical analysis, comparisons were made between two specific groups: (1) sh-NC versus sh-DLAT to evaluate the effects of DLAT knockdown, and (2) sh-DLAT + EV versus sh-DLAT + oeSREBP1 to assess the rescue effects of SREBP1 overexpression. Ns, no significance, ***P < 0.001, and **P < 0.01