Fig. 2.

RCAN1.4 mRNA induction by diverse stimuli in mouse primary mesangial cells. (A) Mesangial cells were isolated from glomeruli by differential sieving, and then the cells of passages 7 to 10 in quiescent state were stimulated with the agents indicated below for 24 h or cultured for 48 h in high glucose condition (33 mM) or in osmotically balanced control medium (5.9 mM glucose plus 27 mM mannitol). Effect of the each stimulation was tested for RCAN1.4 mRNA induction, as described in the Methods. Concentrations were: TGF-β1 (4 ng/ml), ANG II (2 µM), VEGF (10 ng/ml), IL-1β (20 ng/ml), TNF-α (20 ng/ml), MCP-1 (100 ng/ml), oxidative stress (300 µM of H₂O₂), BSA (50 µg/ml), and AGE-BSA (50 µg/ml). ^*p<0.01 compared to no-treatment. (B) Effect of CsA on IL-1β- or AGE-BSA-mediated RCAN1.4 induction was tested. Quiescent mesangial cells were stimulated with IL-1β (20 ng/ml) or AGE-BSA (50 µg/ml) in presence of CsA (1 µM; open bars) or vehicle (filled bars). After 24 h, mRNAs were isolated to determine RCAN1.4 mRNA induction. All the values are means±SE of five experiments/group. Values are expressed relative to the expression observed in untreated controls (arbitrarily assigned a value of 1). ^*p<0.01 vs. the corresponding control (vehicle) group.