Supplementary Figure 5: Cytoskeletal changes and TLR3 trafficking in HSV-1-infected fibroblasts depend on mTOR, PKC and Rab7a.

(a, b) MEF-TLR3-Unc93B1 cells were left untreated (-) or treated with Go 6976 at 250 nM (G) or Torin 1 at 250 nM for 2 h (T). Cells were left uninfected (-) or infected with HSV-1 at an MOI of 10. After 6 h, polymerized α-tubulin and actin were stained (a). The statistical analyses of the mean fluorescence intensity of α-tubulin and actin are shown (b). ***P < 0.001. These experiments were repeated more than three times. (c) MEF-TLR3-Unc93B1 cells were untreated (U) or treated for 2 h with Go 6976 at 250 nM or Torin 1 at 1 μM. The treated cells were left uninfected (U) or infected with HSV-1 at an MOI of 10 for 6 h. TLR3 (green) and nuclei (blue) were stained. The percentages of peripheral TLR3 are also shown. (d) WT and Rab7a^Δ/Δ MEF-TLR3-Unc93B1 cells were left uninfected (U) or infected with HSV-1 at an MOI of 10 for 6 h. TLR3 (green) and nuclei (blue) were stained, and the percentages of peripheral TLR3 were statistically analyzed. Statistical analyses were performed by one-way ANOVA and Tukey’s test. ***P < 0.001, Scale bar, 20 μm. These experiments were repeated more than three times