Fig. 4: Knockdown of TMEM16F in HAP1 cells eliminates Ca2+-activated PS exposure.
a RT-PCR analysis of HAP1 cells (horizon, Cambridge, UK) detected expression of TMEM16D,F,H,K. b Western blots indicating expression of TMEM16D,F,H,K in HAP1 parental cells (Parental) and knockout of expression of TMEM16F (KO_16F) or knockout of TMEM16D,F,H,K (KO_T16all) by gene editing. Very right panel shows loading controls in Parental, KO_16F, and KO-all (β-actin). c, d Dot blot diagram from flow cytometry and summary of annexin V positive cells, indicating Ca2+-dependent activation of phospholipid scrambling (exposure of phosphatidylserine, PS) by ionomycin in parental HAP1 cells but not in cells lacking expression of TMEM16F (KO_16F) or TMEM16F and additional TMEM16 proteins. e Proliferation of HAP1 cells lacking expression of TMEM16F or lacking expression of all endogenous TMEM16 proteins was significantly reduced when compared with parental cells. Mean±SEM (number of experiments). #Significant activation by ionomycin, or inhibition of proliferation (p < 0.05; unpaired t-test)
