Fig 2: Western blotting validation of differentially expressed proteins in mammary tissue between the HV and LV groups. Note: LV: total valine:lysine = 0.63:1; MV: total valine:lysine = 0.73:1; HV: total valine:lysine = 0.93:1. FASN: fatty acid synthase; ACACA: acetyl-CoA carboxylase. All data with error bars represent mean ± SEM. Means not sharing the same letter are different (p < .05)

Fig 3: Lim inhibits the transcriptional activity of SREBP1 and SREBP2 through AMPK in hepatocytes. The AML12 cells were treated with DMSO, 0.4 mM palmitic acid (PA), 0.4 mM PA + 100 µM Lim or 0.4 mM PA + 100 µM Lim +1 µM Comp C for 16 h respectively, after starving in serum-free DMEM for 24 h (A,B) The representative images of SREBP1 and SREBP2 immunofluorescent staining in cells. Scale bar = 300 µm. (C) Quantification of the fluorescent intensity of SREBP1 and SREBP2 in the nuclei relative to that of the cytoplasm. (D,E) The mRNA levels of Fasn, Acc1 and Hmgcr, Hmgcs genes were evaluated using RT-PCR. Values were expressed as mean ± SD (n = 3). * p < 0.05, ** p < 0.01.

Fig 4: Schematic illustration of the antioncogenic effect of CPT mediated by mutant K-Ras protein. KRAS, Kirsten rat sarcoma viral oncogene homolog; CPT, cryptotanshinone; GOT, glutamic-oxaloacetic transaminase; GLUD, glutamate transport system permease protein; GLS, glutaminase; IDH, isocitrate dehydrogenase; FASN, fatty acid synthase; ACC1, acetyl-CoA carboxylase 1; ACLY, ATP-citrate lyase; ROS, oxygen species.

Fig 5: Lipid and carbohydrate metabolism in human hepatoma cell lines, following AKR1D1 knockdown. AKR1D1 knockdown (white bars) increased GLUT1 and GLUT9 and decreased GAPDH and glycogen phosphorylase (PYGL) mRNA expression in HepG2 cells, as measured by qPCR (a). Furthermore, intracellular glycogen storage increased (b), whilst, no differences were observed in total or phosphorylated GSK3ß levels, following 15 min insulin stimulation (5 nM, 50 nM) (c). In addition, AKR1D1 knockdown enhanced lipogenic gene expression (FAS, ACC1, SCD1, SREBF1) (a), reduced the expression of ABCA1 and PPARa (a), and increased total and phosphorylated ACC protein levels (d). Consistent with the lipogenic and PPARa gene expression, AKR1D1 knockdown increased TAG accumulation in HepG2 cells (e) and reduced fatty acid oxidation, as measured by cell media 3-hydroxybutyrate [3OHB] concentrations (f). Moreover, AKR1D1 knockdown increased de novo palmitate synthesis in both TAG and phospholipid fraction, as measured by incorporation of 2H from 2H2O into TAG and phospholipid palmitate, respectively (g). Representative Western blot images are shown from 3 biological replicates, however formal quantification was performed in n = 7–9 replicates. qPCR data were normalised to 18SrRNA and protein data were normalised to ß-tubulin. Data are presented as mean ± se of n = 7–8 experiments, performed in triplicate, *p < 0.05, **p < 0.01, ***p < 0.001, compared to scrambled controls (black bars).