Fig 1: In high Cav-1 cells, Cav-1 mediates LMPt endocytosis, and in low Cav-1 cells, macropinocytosis and Cav-1 induced by LMPt contribute to LMPt endocytosis together. AsPC-1 cells were treated with or without indicated endocytosis inhibitors for 2 h followed by exposure to LMPt for 24 h, LMPt accumulation was observed with fluorescent microscope (A), flow cytometry (B) and ICP-MS (C). Scale bar, 20 μm. ns, no significant; ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, vs. Ctrl. The data are shown as mean ± SD (n = 3). (D) Western blot analysis of endogenous Cav-1 in different pancreatic cancer cells. (E) BxPC-3 cells were transfected with siCav-1 or siNC, cellular accumulation of LMPt was detected with fluorescence microscope. Scale bar, 20 μm. (F) Cells were treated with indicated concentrations of LMPt for 24 h or with 30 μmol/L LMPt for the indicated time, Cav-1 protein was detected by Western blot. (G) AsPC-1 cells were transfected with Cav-1 or vector, and LMPt accumulation was observed with fluorescence microscope, scale bar, 20 μm. (H) AsPC-1 cells were treated with LMPt and siCav-1, then Cav-1 level was detected with Western blot. (I) AsPC-1 cells were treated with siCav-1 in the presence or absence of indicated inhibitors followed by exposure to LMPt. LMPt accumulation was observed with fluorescence microscope. Scale bar, 20 μm.
Fig 2: Mechanism schematic diagram of LMPt involving cellular entry and anti-cancer effect in pancreatic cancer cells. LMPt enters cells via caveolae-mediated endocytosis and macropinocytosis, and Cav-1 level determines the switch of endocytosis pathways. In high Cav-1 cells, only Cav-1 mediates LMPt endocytosis and in low Cav-1 cells, macropinocytosis and LMPt-induced Cav-1 contribute to LMPt endocytosis together. After endosome–lysosome processing, in unchanged metabolite, MPt is released and targets mitochondria to enhance binding of mitochondria protease LONP1 with POLG and TFAM (key players of mtDNA replication) to degrade POLG and TFAM. Then via PINK1–Parkin axis mitophagy is induced based on POLG and TFAM degradation-initiated mtDNA replication block.
Supplier Page from Sino Biological, Inc. for Human Caveolin-1/CAV1 Gene ORF cDNA clone expression plasmid