Fig. 3: Hepatocyte membrane repair is important for invasion of Plasmodium sporozoites.

a Number of infected cells at 2 h after infection, infected with WT (white) or EXP2 cKO sporozoites (blue), after addition of rPfEXP2 at 1 h after infection (N = 5 independent experiments totaling 15 replicates for all concentrations except for 0.5 nM, where N = 3 independent experiments totaling nine replicates were performed). b Number of infected cells at 2 h after infection, infected with WT (white) or EXP2 cKO sporozoites (blue), after addition of rα-HL at 1 h after infection (N = 3 independent experiments totaling nine replicates). c Calcium uptake by cells measured using Gcamp6f fluorescent protein, through time after addition (at time 0 s) of 100 nM rPfEXP2 (blue line) or vehicle control (black line) (N = 3 independent experiments, >75 total cells imaged per condition). d Potassium efflux by cells measured using GEPII fluorescent protein, through time after addition (at time 0 s) of 100 nM rPfEXP2 (blue line), 50 µM of Digitonin (brown line, permeabilizes cellular membranes) or vehicle control (black line) (N = 2 independent experiments, >50 cells imaged per condition). e Number of infected cells at 2 h after infection by GFP-expressing sporozoites in the presence of desipramine (acid SMase inhibitor, white) or GW4869 (neutral SMase inhibitor, gray) (N = 3 independent experiments totaling nine replicates). f Number of infected cells at 2 h after infection by GFP-expressing sporozoites after knockdown of aSMase by shRNA, in the absence (white) or presence of 10 µU/mL of recombinant acid SMase from Bacillus cereus (gray). (N = 3 independent experiments totaling nine replicates). g Number of infected cells at 2 h after infection, infected with WT (white) or EXP2 cKO sporozoites (blue), after treatment with 10 µU/mL of recombinant aSMase added at 1 h after infection. (N = 5 independent experiments totaling 15 replicates). Results are shown as mean±SEM, two-tailed Mann–Whitney U test was applied for p values in all panels except for c, d. c, d Lines represent the average of the mean fluorescence intensity and shaded areas the 95% confidence intervals for each time point. Dose-inhibition curve in e was generated and IC₅₀ value was estimated from the fitted curve.