Fig 2: Protein expression levels of lipid metabolism markers in the liver tissue samples of WT and NLRP3−/− mice. (a) MTTP (b) FASN (c) DGAT1. All data are shown as the means ± SEM (n = 8). ** p ≤ 0.01/*** p ≤ 0.001 (WT Young/Old vs. NLRP3 Young/Old); # p ≤ 0.05/### p ≤ 0.001 (WT Young vs. WT old); ααp ≤ 0.01(NLRP3 Young vs. NLRP3 old).

Fig 3: Blood lipids, hepatic FFA and TG contents, the protein and mRNA expression of liver tissues. (A) Serum TC, TG, HDL-C and LDL-C levels were assessed. (B) Hepatic FFA, TG levels were assessed. (C) Western blot results for CD36, FATP2 in liver samples. (D) Western blot results for ApoB100, MTTP in liver samples. (E-F) ApoB100 and MTTP gene levels of liver samples were measured through qRT-PCR, and normalized against β-actin level. Data were expressed as the means ± SD. *P<0.05, **P<0.01 vs. MN group. #P<0.05, ##P<0.01 vs. HN group. FC, fold change, model-to-normal group ratio. MN, standard chow diet; MCDD, methionine-choline-deficient diet; HN, standard chow diet; HFD, high-fat diet

Fig 4: Lomitapide mesylate and lomitapide inhibit PDAC independently of lipid metabolism, autophagy suppression, and P38 signaling(A) MTTP mRNA expression in human tissues, as retrieved from The Human Protein Atlas database.(B) MTTP mRNA expression in human cancer cell lines, as retrieved from The Human Protein Atlas database.(C) Basal MTTP expression in HepG2, BxPC3, and SW1990 cells.(D) Oil Red O staining of BxPC3 and SW1990 cells treated with 8 μM lomitapide mesylate, or 8 μM lomitapide, or an equivalent volume of DMSO (vehicle control) for 6 h. Scale bars, 200 μm (applies to all images in [D]).(E) LC3B-II and p62 protein expression in BxPC3 and SW1990 cells following the indicated treatments.(F) LC3 transformation assay in cells following treatment with 8 μM lomitapide mesylate, or 8 μM lomitapide, or an equivalent volume of DMSO (vehicle control), in combination with autophagy inhibitors. Cells were pre-treated with autophagy inhibitors (CQ, 20 μM; NH4Cl, 20 mM; or E64D [10 μg/mL] + pepstatin A [10 μg/mL]) for 1 h, followed by treatment with the aforementioned agents for 6 h. Protein extracts were then analyzed for LC3B expression.(G and H) Monitoring autophagic flux in PDAC cells using the mRFP-GFP-LC3 dual-labeling system. BxPC3 and SW1990 cell lines with lentivirus-mediated stable overexpression of stubRFP-sensGFP-LC3 were constructed to track autophagic flux. Following the indicated treatments, the distribution of LC3-positive puncta was visualized via laser confocal microscopy. Yellow fluorescent spots (merged mRFP and GFP signals) represent autophagosomes, while red fluorescent spots (mRFP-only signals, due to GFP quenching in the acidic environment of autolysosomes) indicate autolysosomes. Statistical analysis of the percentages of yellow and red puncta was performed to quantify changes in autophagic flux (H), n = 3. Scale bars, 20 μm (applies to all images in [G]).(I and J) Lomitapide mesylate and lomitapide were added 1 h after pretreatment with autophagy inhibitors or an activator, and cell viability was assessed 6 h thereafter. Autophagy inhibitors and activators used included WM, 5 μM; 3 MA, 5 mM; CQ, 20 μM; NH4Cl, 20 mM; E64D (10 μg/mL) + pepstatin A (10 μg/mL); or rapamycin, 10 μM (n = 3).(K) BxPC3 and SW1990 cells were treated with the indicated treatments for 3 and 6 h, and the target proteins as well as their associated proteins were detected.(L) BxPC3 and SW1990 cells were pre-treated with SB202190 (10 μM) for 1 h, followed by the addition of the indicated treatments; cell viability was then assessed 6 h later (n = 3). Data represent mean ± SD of three independent experiments. Statistical significance was determined using an unpaired two-tailed Student’s t test. ∗, p < 0.05, n.s., not significant.