Journal: Iranian Journal of Basic Medical Sciences
Article Title: Curcumin protects human umbilical vein endothelial cells against high oxidized low density lipoprotein-induced lipotoxicity and modulates autophagy
doi: 10.22038/IJBMS.2021.59969.13297
Figure Lengend Snippet: Curcumin ameliorated autophagy inhibition induced by high ox-LDL in HUVECs through the AMPK/mTOR/p70S6K pathway. A and B: The protein expression levels of LC3-II were measured by Western blotting (n=3). C and D: The protein expression levels of AMPK and p-AMPK were measured by Western blotting (n=3). E and F: The protein expression levels of mTOR and p-mTOR were measured by Western blotting (n=3). G and H: The protein expression levels of p70S6K and p-p70S6K were measured by Western blotting. Expression of these proteins was quantified by densitometry using ImageJ software (n=3). The data are shown as the means±SDs (* P < 0.01 vs. the control group; ** P < 0.01 vs. the control group; # P <0.05 vs. the high ox-LDL group; ## P < 0.01 vs. the high ox-LDL group; & P < 0.05 vs. the compound C group). AMPK, AMP-activated protein kinase; mTOR, mammalian target of rapamycin; p70S6K, p70 ribosomal S6 protein kinase
Article Snippet: Protein extracts were isolated, separated on a 12.5% SDS-PAGE gel and transferred onto a 0.22 μm PVDF membrane (Epizyme), which was incubated with the following primary antibodies: peroxisome proliferator-activated receptor γ (PPARγ) (16001-1-AP, Proteintech, Wuhan, China), interleukin-6 (IL-6) (ab233706, Abcam, Cambridge, 1:1000), interleukin-10 (IL-10) (ab133575, Abcam, 1:5000), tumor necrosis factor alpha (TNF-α) (17590-1-AP, Proteintech, Wuhan, China), microtubule-associated protein 1 light chain 3 (LC3) (2775S, Cell Signaling Technology, MA, USA, 1:1000), AMPK (HN0506, HuaBio, Hangzhou, China, 1:1000), mTOR (HN0824, HuaBio, 1:500), p70S6K (HJ0505, HuaBio, 1:1000), phosphorylated AMPK (p-AMPK) (bs-8813R, Bioss, Beijing, China, 1:1000), phosphorylated mTOR (p-mTOR) (2971S, Cell Signaling Technology, 1:500), and phosphorylated p70S6K (p-p70S6K) (9234S, Cell Signaling Technology, 1:1000). β-Actin (AC026, ABclonal, Wuhan, China, 1:50000) was used as an internal control.
Techniques: Inhibition, Expressing, Western Blot, Software
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