Fig 2: Treatment of T0 and MGF activates AMPK signaling pathway to suppress hepatic lipogenesis and induce autophagy to accelerate hepatic degradation in Apoe−/− mice. A Expression of p-AMPK, AMPK, and CD36 was determined by western blot with total proteins extracted from liver samples of Apoe−/− mice. B Expression of lipid biosynthesis related SREBP-1c, p-ACC, ACC, FAS and SCD-1. C Expression of lipolysis related ATGL, p-HSL and HSL, and β-oxidation relative CPT1. D Expression of autophagy related LC3II, ATG5, ATG7, p-p38 and p38. ***p < 0.001, **p < 0.01, *p < 0.05 (n = 3). E Hepatic red fluorescent LC3 protein was determined by immunofluorescent staining. Scale bar, 50 μm

Fig 3: Cpt1a levels and ceramide in lung tissues from patients with COPD and control group. (A) Western blots and (B) quantification of protein expression for Cpt1a in lung tissues from control and patients with COPD. (C) Ceramide were measured in lung tissues from control and patients with COPD by mass spectrometry. Data are expressed as mean ± standard error of the mean; n=10; **P<0.01 vs. Control; ##P<0.01 vs. COPD with low expression of Cpt1a. Cpt1a, carnitine palmitoyltransferase 1A; COPD, chronic obstructive pulmonary disease.

Fig 4: Increased lipid synthesis is required for hyper-proliferation and survival of human PAH PAVSMC. (A–E) Early passage distal primary human PAVSMC from non-diseased (CTRL) and PAH subjects were serum-deprived for 48 hours and subjected to immunoblot analysis to detect indicated proteins. n = 5 (CTRL), n = 7 (PAH for P-ACLY, ACLY, see Supplementary Figure 1A for additional immunoblots used for statistical analysis) or n = 5 (PAH for ACC, FASN, CPT1A). Data are means ± SE, fold to control. (F,G) Human PAH PAVSMC were incubated for 48 h in serum deprived media supplemented with regular (+ Lipids) or lipid-deprived (-Lipids) 0.1% BSA and subjected to fluorescent BODIPY 493/503 staining (green) to detect intracellular neutral lipids followed by DAPI (blue) staining to detect nuclei. Representative images with enlarged area (F) and statistical analysis (G) are shown. Bar equals 50 μm. Data are means ± SE from n = 5 subjects/group, fold to cells incubated in regular 0.1% BSA (+ Lipids) group. (H) Equal amount of human PAVSMC from five PAH and five control subjects were seeded to 6 well plates. 48 h later (day 0), media was changed to the serum-free media supplemented with regular (+ Lipids) or lipid-deprived (-Lipids) 0.1% BSA; six days later, cell count assay was performed. Data are means ± SE, n = 5 subjects/group. (I) PAH PAVSMC were treated with ACC inhibitor TOFA (20 μM) for 48 h followed by proliferation analysis (Ki-67). Data are means ± SE, percentage of Ki-67 positive cells/total number of cells, n = 5 subjects/group. (J) PAH PAVSMC were treated with ACC inhibitor TOFA (20 μM) for 48 h followed by apoptosis analysis. Data are means ± SE, percentage of TUNEL-positive cells/total number of cells, n = 5 subjects/group. *p < 0.05, **p < 0.01 by Mann–Whitney U test.

Fig 5: CPT1A/Snail axis promotes the development of PAAD(A) qRT-PCR was analyzed the mRNA expression of Snail in PAAD cells.(B) CoIP was detected the interaction between CPT1A and Snail.(C) BxPC-3 cells of vector and CPT1A overexpression groups were treated with 10 μM CHX for 0, 20, 40, and 60 min, then cells were collected and lysed respectively. The half-life of Snail protein was detected by Western blotting and quantified by Prism software.(D) BxPC-3 cells of control and sh-CPT1A groups were pretreated with 10 μM MG132 for 4 h. The ubiquitination levels of Snail were measured by IP experiments.(E and F) MTT (E) and colony formation (F) assays were detected the proliferation ability of CPT1A overexpression cells after knockdown Snail.(G) Transwell assay was detected the longitudinal migration abilities of cells in PAAD cells.(H) Metabolic kits were detected the glucose and ATP levels in PAAD cells.(I) Western blot demonstrated the changes of EMT and glycolysis makers expression after Snail knockdown. ∗p < 0.05 and ∗∗p < 0.01.