Fig. 2

ATP released from activated Müller cells induces microglia activation through P2X7R. a–c Representative immunoblots showing the changes of GFAP and TSPO expression in retinas with intravitreal injections of DHPG (100 μM, 2 μL) (a), DHPG along with Gap26 (200 μM, 2 μL) (b), and DHPG along with BBG (10 μM, 2 μL) (c) at different times after the injections. d, e A comparison of average relative densitometric quantifications of the immunoreactive bands obtained following different kinds of injection is shown in the bar charts (d for GFAP; e for TSPO). f Representative immunoblots showing the changes of GFAP and TSPO expression at different post-operational times in normal and COH retinas with intravitreal injections of saline (2 μL)/20 μM MPEP (2 μL). Saline or MPEP injections were made two days before COH operation. g, h A comparison of average relative densitometric quantifications of the immunoreactive bands obtained following different kinds of injection is shown in the bar charts (g for TSPO; h for GFAP). i Representative immunoblots showing the changes in TSPO and GFAP protein expression in COH retinas with or without the intravitreal injection of BBG. 2 μL BBG (10 μM) or normal saline was intravitreally injected 2 day before the COH operation. j, k A comparison of average relative densitometric quantifications of the immunoreactive bands obtained following different kinds of injection is shown in the bar charts (j for TSPO; k for GFAP). l, m Western blot results showing the changes in TSPO protein expression in COH retinas with or without the intravitreal injection of 5-BDBD, a P2X4R antagonist. All the data are normalized to control. In all experiments, n = 6 for each group. *P < 0.05, **P < 0.01, and ***P < 0.001 vs. Ctr using ordinary one-way ANOVA with Dunnett’s multiple comparisons test (d, e, g, h, j, k, m); ^#P < 0.05 and ^###P < 0.001 vs. corresponding saline group at the same time point using paired two-tailed Student’s t test (g, h, j, k)