Figure 1: Effect of siRNA to PKCα on calcium uptake in cultured intestinal cells. Isolated cells were resuspended in 40 ml of GBSS (Gey’s balanced salt solution) and 200 μl pipetted into 35 mm plastic Petri dishes containing 3 ml of RPMI-1640 medium and antibiotics (100 units/ml penicillin, 100 mg/ml streptomycin, Sigma). The cells were incubated for 24 h (in the absence of serum) at 37°C with 5% CO2/95% air to promote cell adhesion and then transfected. The transfection protocol is described in material and methods. After 24 h cultured cells were incubated with or without 300 pM 1,25(OH)2D3 in 1 ml of GBSS/0.1% (wt/vol) BSA containing 4 μCi of 45CaCl2 for 7 min. The cells were rinsed 3 times with 4 ml ice-cold GBSS, followed by the addition of 0.5 ml of 0.1% (vol/vol) Triton X-100 in 10 mM TED to each plate. Cells were scraped and radioactivity determined by liquid scintillation counting. Data are represented as mean ± S.E.M. (***P < 0.001, relative to corresponding controls).