FIG 2.
DPM1 and DPM3 trans-complementation restores cell susceptibility to DENV infection. (A) Immunoblot of DPM1 in control, DPM1KO, and DPM3KO cells trans-complemented with the respective cDNA. (B) Cell surface CD59 staining on 293T and HAP1 clones trans-complemented with DPM1 or DPM3 cDNA. (C) 293T and HAP1 trans-complemented clones were challenged with DENV2 JAM (MOI of 5 in 293T cells and MOI of 10 in HAP1 cells). Infection was quantified 48 hpi by flow cytometry using MAb 2H2. Data are means ± SD from three independent experiments performed in duplicate. Significance was calculated using a one-way ANOVA with Dunnett’s multiple-comparison test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.001. (D) Control and DPM1 and -3 KO Huh-7 clones were inoculated with increasing MOIs of DENV2 JAM. Levels of infection were quantified 48 hpi by flow cytometry using MAb 2H2. (E) HFF1 cells transfected with control, DPM1, or DPM3 sgRNA were inoculated with increasing MOIs of DENV2 JAM. Levels of infection were quantified 48 hpi by flow cytometry using MAb 2H2 in aCD59-negative population. (A, B, D, and E) Data are representative of results from at least two independent experiments.