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. 2017 Mar 29;91(8):e01564-16. doi: 10.1128/JVI.01564-16

FIG 2.

FIG 2

TMPRSS2 activity directly mediates bafilomycin A-independent MHV infection. (A) JHM.SD is more sensitive than A59 to TMPRSS2 transfection. HEK-293T cells cotransfected with pCAGGS-mCeacam1a-4L and pCAGGS-hTMPRSS2-FLAG were infected with the indicated virus. Using two-way ANOVA (n = 5), P values were <0.0001 for the effects of TMPRSS2 and the virus strain and their interaction. Asterisks indicate the TMPRSS2 transfection levels at which the 2 viruses were significantly different from each other by Tukey's multiple comparison. (B) Camostat abrogates the effect of TMPRSS2 on JHM.SD infection. Transfected HEK-293T cells were treated with DMSO or camostat (final DMSO concentration, 1.5%) prior to infection. Using two-way ANOVA, P values were <0.0001 for TMPRSS2 transfection, camostat treatment, and their interaction. Number signs indicate significant differences from the no-TMPRSS2 control within the DMSO group using Dunnett's multiple comparisons (no significant difference from the no-TMPRSS2 control at any level of TMPRSS2 transfection within the camostat group). (C) TMPRSS2 enhances JHM.SD infection in the presence of bafilomycin A. Transfected HEK-293T cells were treated with DMSO or bafilomycin A (final DMSO concentration, 0.5%) prior to infection with JHM.SD-fluc. Using two-way ANOVA, P values were <0.0001 for the effects of TMPRSS2 and bafilomycin A and their interaction. Symbols: #, significant difference from the no-TMPRSS2 control within the DMSO group; †, significant difference from the no-TMPRSS2 control within the bafilomycin A group (determined by Dunnett's multiple-comparison test). (D) TMPRSS2 overcomes bafilomycin A inhibition of A59 infection. Transfected HEK-293T cells were treated with DMSO or bafilomycin A (final DMSO concentration, 0.5%) prior to infection with rA59-fluc. Using two-way ANOVA, P values were <0.0001 for the effects of TMPRSS2 and bafilomycin A and their interaction. Symbols: #, the TMPRSS2 transfection levels at which A59 infection differed from the no-TMPRSS2 control within the DMSO group; †, the TMPRSS2 transfection levels at which A59 infection differed from the no-TMPRSS2 control within the bafilomycin A group (determined by Dunnett's multiple-comparison test). All data are representative of at least 2 independent experiments with n = 5 technical replicates.