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[Preprint]. 2023 Dec 17:2023.12.17.572063. [Version 1] doi: 10.1101/2023.12.17.572063

Figure 5. Knockdown of EHD1 or MICAL-L1 does not reduce cell surface Env but diminishes specific particle infectivity.

Figure 5.

(A) Knockdown or Scr cells were infected or transfected as described in the methods for Figure 5 and stained for cell surface Env with APC-conjugated 2G12 antibody. Cells were then permeabilized and stained for p24 with KC-57 FITC to gate for infected cells. Neither EHD1 nor MICAL-L1 knockdown (blue) reduced cell surface Env relative to Scr control (red) in any of the cell types tested. Grey indicates uninfected control cells stained with 2G12 antibody. (B) Virus was produced from EHD1 or MICAL-L1 knockdown cells as in experiments shown in Figure 4 and evaluated for specific infectivity by TZM-bl assay. EHD1 knockdown is shown in light colored bars, and MICAL-L1 knockdown is shown in the darkest colored bars. Semi-permissive HeLa and permissive 293T and MT4 cells showed no significant difference in particle infectivity from Scr treated or knockdown cells. In nonpermissive H9, CEM, and Jurkat cells there was a significant reduction in infectivity of particles produced following MICAL-L1 or EHD1 depletion. Significance was assessed using ANOVA with false discovery rate correction as before. *, P<0.05; **, P<0.01; ***, P<0.001.