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Fig. 4 | Journal of Neuroinflammation

Fig. 4

From: P2X7R-mediated autophagic impairment contributes to central sensitization in a chronic migraine model with recurrent nitroglycerin stimulation in mice

Fig. 4

Activation of autophagy reduced NTG-induced CGRP and c-fos expression in the TNC. a–c Representative western blots showing the change of autophagy-related proteins after RAPA treatment. Quantification of band intensities revealed increased ration of LC3II/LC3I (a), in combination with decreased p62 (b), as well as upregulated beclin1 (c) after chronic RAPA and NTG administration compared with NTG administration only. d Immunoblots of P2X7R showed that autophagy activation did not affect the expression of P2X7R. e Representative immunoblots of CGRP and c-fos after RAPA intervention. The optical density analysis indicated RAPA treatment significantly decreased the expression of CGRP and c-fos compared with NTG administration only. n = 6 per group; data are presented as mean ± SEM; one-way ANOVA and Tukey’s multiple comparison test; **p < 0.01 and ***p < 0.001 compared with the SHAM-VEH group; #p < 0.05 and ###p < 0.001 compared with the NTG-VEH group. f The white dotted line frame indicated the representative image of the TNC area used for the analyses. g, j Immunofluorescence images of CGRP (g) and c-fos (j) in the TNC area. Scale bars: 100 μm for CGRP, 20 μm for c-fos. h, i The CGRP immunoreactivity (h) and c-fos-positive cells (i) were reduced by RAPA treatment. n = 4 per group; 6 sections from each mouse were analyzed; values are expressed as mean ± SEM; one-way ANOVA and Tukey’s multiple comparison test; ***p < 0.001 compared with the SHAM-VEH group for CGRP or the SHAM group for c-fos ; ###p < 0.001 compared with the NTG-VEH group for CGRP or the NTG group for c-fos

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