Syk and Hrs Regulate TLR3-Mediated Antiviral Response in Murine Astrocytes

Figure 8

Poly(I:C) treatment of murine astrocytes induces TLR3 tyrosine phosphorylation and promotes interaction with Hrs. (a) After poly(I:C) or poly(I:C)/LyoVec stimulation for 5, 8, 12, 15, and 30 min, C8-D1A cells were lysed and TLR3 was immunoprecipitated using the anti-TLR3 antibody. Phosphotyrosine (P-Tyr), Hrs, and STAM were then detected by Western blot. (b) Following poly(I:C) or poly(I:C)/LyoVec stimulation for 5, 8, 12, andn 15 min, C8-D1A cells were lysed and TLR3 was immunoprecipitated using the anti-TLR3 antibody. Ubiquitin was detected by Western blot. Blue arrows indicate ubiquitinated TLR3. (c) Following poly(I:C) stimulation for 5, 8, 12, and 15 min, murine astrocytes were lysed and STAM and Hrs were immunoprecipitated using anti-STAM and anti-Hrs antibodies, respectively. Hrs and STAM were detected by Western blot. For all immunoprecipitation experiments, 0 min presents untreated cells and mouse IgG were used as a negative control. EL: immunoprecipitation eluate; FT: immunoprecipitation flow through; CN: control cell lysate. GAPDH was used as protein loading control.

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