Figure 5.

Effect of in vivo CD200R1 and miR-129-5p inhibition on the level of microglial TNF-α and IL-6. Animals were exposed to arsenic (0.38 mg/kg bd. wt.) for 2 months, and in the last week of exposure, animals were treated with CD200R1 siRNA and anti-miR-129 intracerebrally by stereotaxic method and sacrificed on 60^th day. A, experimental scheme, (B) exposure to arsenic increased the level of miR-129-5p, which was brought down by the treatment of anti-miR-129 (n = 3 mice/group). CD200R1 siRNA did not show any significant effect on the level of miR-129-5p. C, arsenic exposure did not alter the mRNA level of CD200R1 like in vitro compared with control, whereas siRNA significantly inhibited it. D, Western blot analysis showed that arsenic exposure inhibited CD200R1 protein, which was rescued by anti-miR (n = 5 mice/group). E, immunofluorescence staining of CD200R1 in brain sections (n = 3 mice/group). Scale bar: 50 μm for uncropped images and 17 μm for the cropped images. The effects of CD200R1 siRNA and anti-miR-129 on the level of (F) TNF-α (n = 3–4 mice/group) and (G) IL-6 (n = 5 mice/group) were measured in the ex vivo microglial culture supernatant. Bar graphs represent mean ± SEM. “p” denotes the level of significance in comparison to control; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001; ns, nonsignificant.