Figure 5.

RGS12 promotes the polarization of macrophages toward the M1 phenotype. (A, B) RAW264.7 cells were transfected with pCMV-empty or pCMV-RGS12 plasmids for 48 h, and the M1 macrophage marker CD86 and M2 macrophage marker CD163 were detected by the immunofluorescence staining assay. (A) RAW264.7 cells were incubated with the anti-CD86 or anti-CD163 antibody followed by incubation with fluorescein isothiocyanate–conjugated secondary antibody, and the nuclei were stained with DAPI (blue). Images show CD86 in red and CD163 in green. (B) The expression of CD86 and CD163 in each cell was analyzed by the ImageJ densitometry method. Ten views in each well were analyzed. The relative intensity of the fluorescence was determined by comparing each intensity value to the average intensity. The blue bar represents pCMV-empty (Ctrl) and the red bar represents the pCMV-RGS12 overexpression (RGS12 OE) (bar, 10 µm). n = 5, ***P < 0.001. (C–E) RAW264.7 cells were transfected with pCMV-RGS12 or pCMV-empty (Ctrl) plasmids for 48 h, and the M1 macrophage markers were detected by real-time polymerase chain reaction (PCR) assay. Note that overexpression of RGS12 can increase the IL1β, IL6, and TNFα levels. **P < 0.01, ***P < 0.001 versus the control group, n = 5. (F–H) RAW264.7 cells were treated as described in (C) to (E), and the M2 macrophage markers (IL4, IL1RA, and IL10) were detected by real-time PCR assay. Values are means ± SEMs (P > 0.05 versus the control group, n = 5). (I) Schematic for RGS12 role in periodontitis. RGS12 promotes the polarization of M1 macrophages, the migration of macrophages, and the expression of proinflammatory factors. Moreover, RGS12 enhances osteoclast formation under inflammatory conditions. Finally, the conditional knockout of RGS12 prevents inflammation and alveolar bone erosion during periodontitis.