Figure 11. Effects of RNAi for P2Y₁ mRNA on Ca²⁺-mobilization and GC secretion in H295R.

shRNA plasmid-transfected H295R were cultured on coverslips or in 24-well plates for 48 hr. The cells were also loaded with fura2-AM for calcium assay. A: Image of eGFP signal-expressing H295R cultured on coverslips (excitation: 488 nm and elimination: 510 nm), B: The same field image as (A) but under the (excitation: 380 nm and the elimination at 510 nm. C: [Ca²⁺]_i traces in cell-1 to cell-4 indicated in (A) and (B). Each scale bar indicates 50 µm. D: Effect of shRNA transfection on 2MeS-ATP-induced glucocorticoid secretion in H295R. Cells were co-transfected with both the shRNA plasmid for P2Y₁ and the eGFP plasmid (KD.). In control H295R (Ctl.), only the eGFP plasmid was transfected. GC secretion by 2MeS-ATP (106.4±5.27) was reduced in shRNA/eGFP-transfected H295R (52.6±6.49). Data represent the Mean±SE (N = 4). The ‘***’ indicates statistical significance at p<0.001. E: Western blotting analysis. I: P2Y₁ in Ctl., II: P2Y₁ in KD., III: GAPDH in Ctl., IV: GAPDH in KD.