Fig. 3

Expression profile of P2X4R after chronic intermittent administration of NTG. a, b Western blot assay for the protein expression of P2X4R on different days. One-way ANOVA and Tukey’s multiple comparison test. n = 5/group, **p < 0.01, ***p < 0.001, versus the vehicle group (VEH). c qRT-PCR analysis for the mRNA expression of P2X4R between the vehicle group and the NTG 9d group (NTG intermittent injection for 9 days). Unpaired t test, n = 6/group. ***p < 0.001. d, e Double immunofluorescence labeling of P2X4R (green) and Iba1 (red) in VEH and NTG 9d groups. Most P2X4R-positive cells are double-labeled (yellow) with Iba1. Scale bars, 20 μm