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. 2024 Oct 25;166(6):1296–1313. doi: 10.1097/j.pain.0000000000003460

Figure 6.

Figure 6.

Validation of myeloid-cell-specific TGR5 knockdown and the effect of TGR5 knockdown on mechanical allodynia. (A) Schematic of experimental design. Representative flow cytometry pseudocolor dot plots indicating the percentage of CD11b+ cells among all GFP+ cells (B) and the percentage of GFP+ cells among all CD11b+ cells (C) in the sciatic nerve of mice 5 days after virus injection. (D) mRNA expression of Tgr5 in the sciatic nerve of Lyz2-Cre mice treated with shControl virus or shGPBAR1 virus. n = 6 per group. *P < 0.05; unpaired t test. (E) Representative Western blotting bands and (F) densitometric quantification of TGR5 in the sciatic nerve of Lyz2-Cre mice treated with shControl virus or shGPBAR1 virus after pSNL. n = 5 per group. ***P < 0.001; unpaired t test. (G) Representative microphotographs of immunofluorescence staining of TGR5 in the sciatic nerve of Lyz2-Cre mice treated with shControl virus or shGPBAR1 virus. Scale bar: 100 μm. (H) Analysis of the TGR5+ mean fluorescence density. n = 4 per group. *P < 0.05; unpaired t test. (I) Comparisons of the 50% paw withdrawal threshold evaluated by the Von Frey test at POD7 between the shControl and the shGPBAR1 group. n = 8 to 9 per group. *P < 0.05; 2-way ANOVA, Šídák post hoc test, F (2, 45) = 6.523, F (2, 45) = 157.7, F (1, 45) = 4.323. Data are presented as the mean ± SEM. ANOVA, analysis of variance; POD, postoperative day; pSNL, partial sciatic nerve ligation.