Figure 2
Augmented SMAD dependent TGFβ signaling in HPASMCs on exposure to combined treatment of cocaine and HIV-Tat. Quiescent HPASMCs were treated with cocaine 1 µM and/or Tat (25 ng/ml) for 2 days for western blot and RNA analysis. (a) Representative images of the Western blots probed using total and phosphorylated (p) - SMAD2/3. Graph represents average densitometry of 3 independent experiments. (b) Real time RT-PCR analysis of PAI1 mRNA expression. (c) HPASMC (3 × 103/well) were seeded in 96 well plate followed by either transfection with siRNASMAD2/3 or siRNAscrambled. After 48 h, the medium was replaced with 0.1% serum containing medium followed by cocaine and/or Tat treatment for 2 days. MTS cell proliferation assay was then conducted. All values are mean ± SD of at least three independent experiments performed in triplicates. *p ≤ 0.05 **p ≤ 0.01, ***p ≤ 0.001 compared to untreated control (Cont.), $p ≤ 0.05, $$$p ≤ 0.001 compared to Tat; #p ≤ 0.05, ###p ≤ 0.001 compared to cocaine (Coc), @p ≤ 0.001 compared to C + T.
