treatment with vehicle dmso  (TargetMol)

Journal: International journal of molecular sciences

Article Title: TMAO Suppresses Megalin Expression and Albumin Uptake in Human Proximal Tubular Cells Via PI3K and ERK Signaling.

doi: 10.3390/ijms23168856

Figure Lengend Snippet: Figure 6. PI3K and ERK mediate TMAO-induced suppression of megalin expression. HK-2 cells were pre-incubated with DMSO (vehicle), Akt inhibitor MK-2206 (1 µM), mTOR inhibitor ridaforolimus (1 µM), PI3K inhibitor wortmannin (1 µM) or ERK inhibitor PD98059 (1 µM) for 1 h prior to TMAO stimulation (300 µM) for 24 h (A) followed by evaluating megalin protein expression with flow cytometry. Megalin protein expression is presented as % of vehicle control (DMSO or respective inhibitor alone), which is represented by the dotted line. Western blot analysis was conducted to identify differences in protein levels of p-Akt/Akt, p-mTOR/mTOR, p-ERK/ERK and PI3K p- P85/PI3K P85 after TMAO (300 µM) stimulation for 3, 5, 15 and 30 min (B,C). GAPDH was used as a loading control. Data are presented as mean ± SEM (n = 3 independent experiments). MFI, mean fluorescence intensity. Asterisks denote statistical significance compared to unstimulated cells (* p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: The HK-2 cells were also treated with TMAO (300 μM) for 12 h followed by treatment with DMSO (vehicle), candesartan (10 μM, Santa Cruz Biotechnology), dapagliflozin (10 μM, Santa Cruz Biotechnology), losartan (10 μM, Santa Cruz Biotechnology) and enalaprilat (5 μM, Santa Cruz Biotechnology Inc) for an additional 12 h in the presence of TMAO.

Techniques: Expressing, Incubation, Cytometry, Control, Western Blot