Figure 1

Homocysteine induces MIP- 2 mRNA (A) and Protein (B) in mesangial cells. MCs were incubated with L-cysteine (100 μM) or Hcy (15 μM, 50 μM and 100 μM) for 24 hours at 37°C in 100 mm dish. To determine expression (A), following trypsinization of cell monolayers, total RNA was isolated by the single-step method [19]. Subsequently, qRT-PCR was performed as described in text. Total protein was extracted from harvested cells under non-denaturing conditions using lysis buffer. MIP-2 protein levels (B) were detected by western blot. Results are representative of three separate experiments. Protein bands were quantified (Quantity One software, Bio-rad) and levels were represented as percentage response of control (100 μM L-Cysteine). Data represent mean ± SEM from three separate experiments. *p < 0.05.