Fig. 3: ATP9A-knockdown cells are neomycin sensitive and expose PI4P in the plasma membrane extracellular leaflet.
From: P4-ATPases control phosphoinositide membrane asymmetry and neomycin resistance
a, Neomycin sensitivity of WT HeLa cells and ATP9A-knockdown cells. The data represent the per cent of viable cells relative to the viable cells in absence of the neomycin. NS, non-silencing (control) siRNA. A mixed effect model was performed to test the variance and comparisons with control cells were made with Dunnett’s multiple comparisons test (n = 3 biologically independent experiments; **P = 0.0054, 0.0025). The data are the mean ± s.d. b, ATP9A-knockdown HeLa cells expose PI4P in extracellular leaflets. Hoechst dye was used as nuclear stain. Scale bar, 10 μm. c, A quantification of GFP–SidCP4C and GFP–PLCPH binding to the cell surface of ATP9A-knockdown cells. A two-way ANOVA was performed to test the variance and comparisons with control siRNA treated cells were made with Dunnett’s multiple comparisons test (n = 20 cells from three biologically independent experiments; ****P < 0.0001). The data are the mean ± s.d. d, Neomycin sensitivity of WT HEK293 cells and ATP9A-knockdown cells. The data represent the per cent of viable cells relative to the viable cells in absence of the neomycin. A mixed effect model was performed to test the variance and comparisons with control cells were made with Dunnett’s multiple comparisons test (n = 3 biologically independent experiments; **P = 0.0024, 0.0014). The data are the mean ± s.d. e, ATP9A-knockdown HEK293 cells expose PI4P in extracellular leaflets. Scale bar, 10 μm. f, A quantification of GFP–SidCP4C and GFP–PLCPH binding to the cell surface of ATP9A-knockdown cells. A two-way ANOVA was performed to test the variance and comparisons with control siRNA treated cells were made with Dunnett’s multiple comparisons test (n = 20 cells from three biologically independent experiments; ****P < 0.0001). The data are the mean ± s.d.
