Supplementary Figure 3: Cytoskeletal changes and TLR3 trafficking are mediated by PKC and mTORC2.
From: Combating herpesvirus encephalitis by potentiating a TLR3–mTORC2 axis
(a, b) NIH3T3-TLR3-Unc93B1 cells were left untreated or treated with Rapamycin at 400 nM (R), Torin 1 at 250 nM (T), or Go 6976 at 250 nM (G) for 2 h. Cells were left unstimulated or stimulated (PIC) with poly(I:C) at 25 μg/ml for 18 h. Polymerized α-tubulin and actin were stained. The mean fluorescence intensity of α-tubulin and actin was statistically analyzed (b). Statistical analysis was performed by one-way ANOVA and Tukey’s test. ***P < 0.001. Scale bar, 20 μm. These experiments were repeated more than three times. (c) The wildtype and RictorΔ/Δ fibroblasts were transduced to express TLR3 and Unc93B1. These cells were left unstimulated (U), stimulated (PIC) with poly(I:C) at 25 μg/ml for 16 h or infected (HSV-1) with HSV-1 at an MOI of 10 for 6 h. TLR3 (green), LAMP-1 (red) and nuclei (blue) were stained and analyzed by confocal microscopy. The statistical analyses of the peripheral TLR3 is also shown. Statistical analysis was performed by one-way ANOVA and Tukey’s test. ***P < 0.001, Scale bar, 20 μm. These experiments were repeated more than three times
