Figure 3

Hcy increases phosphorylation of p38MAPK (A) and p85 PI3 kinase (B). Mesangial cells were serum-starved overnight prior to exposure to medium containing L-cysteine (100 μM) or Hcy (15 μM, 50 μM and 100 μM). Cells were washed with PBS (4°C) and harvested using lysis buffer under non-denaturing conditions. Total p38 MAPK, total p85 PI-3K, phosphorylated p38 MAPK and phosphorylated p85 PI-3 Kinase expression was detected by western blot as described in methods. Protein bands were quantified and the ratios of pp38MAPK/p38MAPK and pp85/p85 were represented as fold-changes compared to t=0. Panel depict representative blot of three separate experiments performed in duplicates; values are expressed as mean ± SEM; *p < 0.02; #p < 0.05.