FIG 7.

Innate-immunity activation is higher upon infection of cells knocked down for p62 or OPTN. (A) Development of p62 knockdown and OPTN knockdown HEL cells with the aid of lentiviral vectors. (B) HEL cells or the p62 knockdown or OPTN knockdown derivatives were infected with either HSV-1(F) or the ΔICP0 virus (0.01 PFU/cell), and cells were collected at 3, 24, and 48 h after infection. Titration of progeny virus was then done in Vero cells for the wild-type virus or in U2OS cells for the ΔICP0 virus. (C) HEL cells or their p62 knockdown derivatives were uninfected or infected with the D199A virus (0.01 PFU/cell). Cells were harvested at 12 h postinfection, and qPCR analysis was done using primer pairs against ISG56 or the 18S rRNA, which served as a normalization control. (D) HEL cells or their p62 knockdown derivatives were uninfected or infected with the HSV-1 Δγ₁34.5 virus (0.01 PFU/cell). Quantification of ISG56 transcripts was done as described above. (E) HEL cells or their OPTN knockdown derivatives were uninfected or infected with the HSV-1 D199A, Δγ₁34.5, or ΔUL46 mutant virus (0.01 PFU/cell). Cells were harvested at 12 h postinfection, and qPCR analysis was done using primers against ISG56 transcripts or the 18S rRNA, as described above. All experiments were repeated three times, and representative results are depicted.