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The autophagy-inducing effects of MK-2206 (MK) and triciribine (TCN), along with their impact on LDLR and ABCA1 expression, were evaluated in HepG2 cells. HepG2 cells were cultured for 14-16 h in the absence (DMSO) or the presence of A) MK-2206 (5 µM) or B) triciribine (0.5 µM) alone, or C-D) pretreated with SBI-0206965 (SBI; 6.4 µM) for 30 minutes prior to administration. Protein levels of ABCA1, LDLR, LC3BI and LC3BII were analyzed by immunoblotting, showing one representative western blot. Quantified values were normalized to β-actin and expressed relative to levels observed in untreated cells (n = 4). Quantitative PCR was used to assess gene expression of ABCA1 , LDLR , LC3B , ULK1 and p62 , with values normalized to GAPDH and expressed relative to levels observed in untreated cells (n = 3). Error bars represent SD. * P < 0.05, ** P < 0.01, and *** P < 0.001 compared with untreated cells.

Journal: PLOS One

Article Title: Autophagy regulates MK-2206-induced LDL receptor expression and cholesterol efflux pathways

doi: 10.1371/journal.pone.0338076

Figure Lengend Snippet: The autophagy-inducing effects of MK-2206 (MK) and triciribine (TCN), along with their impact on LDLR and ABCA1 expression, were evaluated in HepG2 cells. HepG2 cells were cultured for 14-16 h in the absence (DMSO) or the presence of A) MK-2206 (5 µM) or B) triciribine (0.5 µM) alone, or C-D) pretreated with SBI-0206965 (SBI; 6.4 µM) for 30 minutes prior to administration. Protein levels of ABCA1, LDLR, LC3BI and LC3BII were analyzed by immunoblotting, showing one representative western blot. Quantified values were normalized to β-actin and expressed relative to levels observed in untreated cells (n = 4). Quantitative PCR was used to assess gene expression of ABCA1 , LDLR , LC3B , ULK1 and p62 , with values normalized to GAPDH and expressed relative to levels observed in untreated cells (n = 3). Error bars represent SD. * P < 0.05, ** P < 0.01, and *** P < 0.001 compared with untreated cells.

Article Snippet: MK-2206–2HCl (MK-2206; S1078) and triciribine (TCN; S1117) from Selleck Chemicals (Houston, TZ), and Rapamycin (R8781) and SBI-0206965 (SBI; SML1540) from Sigma-Aldrich were utilized as indicated in the figure legends.

Techniques: Expressing, Cell Culture, Western Blot, Real-time Polymerase Chain Reaction, Gene Expression