Figure 4. Diminished ATP-induced ROS production due to depletion of P2X4 or P2X7 by RNA interference.
(A) Immortalized GEC were transduced with lentiviruses carrying the indicated shRNA-expressing plasmid for 1 day and selected with puromycin. After selection, cells were collected and total RNA was analyzed by qPCR to confirm knockdown efficiency. (B) DCF staining of ROS production after 3 mM ATP stimulation for 1 hour in different cell lines. The fluorescence shown in (C) was quantified as in Figure 1 and normalized to shCtrl, which was transduced with control, non-mammalian shRNA.