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. 2019 Sep 17;20:554–566. doi: 10.1016/j.isci.2019.09.020

Figure 4.

Figure 4

BRCA1 Is a Downstream Target of C286 in NP Modulation

(A) Schematic of experimental paradigm.

(B and C) (B) Measurement of thermal and mechanical sensitivities by von Frey filaments (C) and hot plate in vehicle (n = 6) or C286 (n = 6). Data shown as Mean ± SEM. Two-way ANOVA with Pairwise Multiple Comparison Procedures (Holm-Sidak method). ***p ≤ 0.001.

(D) Co-labelling of GFP and Iba1 (scale bar, 20 μm), and GFP and BRCA1 in LVsc + C286- and LVBRCA1shRNA + C286-treated rats (scale bar, 100 μm).

(E–H) (E and F) Western blots and quantification of BRCA1 in L5-SNL rats transduced with LV/SC + C286 (n = 3) or LV/BRCA1shRNA + C286 (n = 3). Student's t test. *p ≤ 0.05 (G) Immunohistological confirmation of LV transduction (scale bar, 20 μm), (H) higher-magnification inset shows BRCA1 in microglia in an LV/Sc + C286-treated rat (scale bar, 10 μm).

(I) Quantification of BRCA1 in microglia.

(J and K) Expression and quantification of γH2AX (scale bar, 20 μm), (L) higher-magnification inset shows γH2AX in microglia in an LV/BRCA1shRNA + C286-treated rat (scale bar, 10 μm).

(M and N) Expression and quantification of CGRP in laminae I–III of the dorsal horn (scale bar, 100 μm).

(O and P) Expression and quantification of P2X4R (scale bar, 100 μm). Inset shows higher-magnification image (scale bar, 20 μm) Data are shown as Mean +SEM of FI. Student's t test, **p ≤ 0.01, ***p ≤ 0.001 (n = 3 per group, 5 sections per animal).