Figure 2.
Tlr8 knockdown promotes dendritic growth in vitro and in vivo. (A) Immunoblotting using HA tag antibody to examine the effect of TLR8 knockdown constructs on TLR8 expression in HEK293T cells. Two expression vectors were used to express miR-Ctrl and miR-Tlr8 as indicated. (B) Tlr7−/−-cultured neurons were transfected with miR-Ctrl and miR-Tlr8 at 2 DIV, and then treated with CL075/poly dT at 4 DIV. Dendritic morphology was analyzed at 5 DIV based on GFP signals. The sample sizes (n) of the examined neurons are indicated. Data were collected from two independent experiments. (C and D) IUE with miR-Ctrl or miR-Tlr8 was performed at E15.5 with WT (C) and Tlr7−/− (D) mice. Layer 2/3 cortical neurons were outlined by GFP signal at P7, P14, and P21 as indicated. N, number of examined mice; n, number of examined neurons. Bars: (B) 20 µm; (C and D) 50 µm. The data are presented as mean + SEM (error bars). *, P < 0.05; **, P < 0.01; ***, P < 0.001. Two-way ANOVA with Bonferroni’s multiple comparisons test (B) and two-tailed nonparametric Mann–Whitney tests (C and D) were used.