Fig. 1.

Oxidized low-density lipoprotein (oxLDL) induces vimentin secretion in 3T3-L1-derived adipocytes. (A) 3T3-L1-derived adipocytes were treated with or without oxLDL (50 µg/mL) for 8, 16, 24 hours and the concentrations of extracellular vimentin in the media were measured using enzyme-linked immunosorbent assay (ELISA). Data were normalized to the protein amount (mg) of the adipocytes used in the assay (n=3). (B) Detection of vimentin in the concentrated media of 3T3-L1-adipocytes cultured with LDL (50 µg/mL) or different concentrations of oxLDL (25, 50 µg/mL) for 24 hours. In the Western blot, His (molecular weight, 1.6 kDa)-tagged recombinant vimentin was used for a positive control. Data were normalized to the protein amount (mg) of the adipocytes obtained from the individual culture dish (n=3). (C) Detection of vimentin in the concentrated media of 3T3-L1-adipocytes cultured with LDL (50 µg/mL for 24 hours) or oxLDL (50 µg/mL for 8, 16, and 24 hours) using Western blotting. Data were normalized to the cellular protein (mg) (n=3). (D) 3T3-L1-adipocytes were pretreated with anti-CD36 antibody (2 µg/mL) before incubating with LDL (50 µg/mL for 24 hours) or oxLDL (50 µg/mL for 24 hours). Detection of vimentin in the concentrated media of the adipocytes using Western blotting and data were normalized to the cellular protein (mg) (n=3). (E) 3T3-L1-adipocytes were pretreated with or without H-89 (10 µM) before incubating with LDL (50 µg/mL for 24 hours) or oxLDL (50 µg/mL for 24 hours). Detection of vimentin in the concentrated media of adipocytes using Western blotting and data were normalized to the cellular protein (mg) (n=3). All experiments were done more than three times. One-way analysis of variance (ANOVA) with Bonferroni post hoc test was done. SF, serum free. ^aP<0.05, ^bP<0.01, ^cP<0.001.