Fig. 5
From: TRPA1 exacerbates selective retinal ganglion cell vulnerability under acute ocular hypertension
Effect of TRPA1 activation on the retinal expression of Ca(2+)/CaMKII/CREB signaling pathways under AOH. (A-C) The mRNA level of (A) Trpa1, (B) Camk2a and (C) Camk2b in retina at day 7 in AOH eyes without or with conditional knock out of Trpa1 in RGCs compared with the contralateral control eyes (n = 4 for each group, unpaired t-test, **P < 0.02, ***P < 0.01). (D) Western blot and the relative optical densities analysis of the level of TRPA1 in (F)Trpa1fl/fl;Thy1-Cre− group and (G)Trpa1fl/fl;Thy1-Cre+ group without or with AOH (n = 4 for Trpa1fl/fl;Thy1-Cre− group, n = 3 for Trpa1fl/fl;Thy1-Cre+ group, paired t-test, *P < 0.05). (H) The relative phosphorylation of CaMKII (calculated as p-CaMKII/total CaMKII) in Trpa1fl/fl;Thy1-Cre− group and Trpa1fl/fl;Thy1-Cre+ group (n = 4 for Trpa1fl/fl;Thy1-Cre− group, n = 3 for Trpa1fl/fl;Thy1-Cre+ group, unpaired t-test, *P < 0.05). (E) Western blot and (I) the relative level of CREB1 phosphorylation (n = 4 for Trpa1fl/fl;Thy1-Cre− group, n = 3 for Trpa1fl/fl;Thy1-Cre+ group, unpaired t-test, *P < 0.05). (J) Immunofluorescent staining of the inner retina (GCL + IPL + INL), showing co-localization and expression of CaMKII and p-CaMKII, and (K) CREB1 and p-CREB1. (L) Western blot and the optical densities analysis of the relative level of (M) CaMKII, (N) CaMKII phosphorylation and (O) CREB1 phosphorylation (n = 3 for each group, unpaired t-test, **P < 0.02, ***P < 0.01). GAPDH was used as the internal controls. All data were presented as mean ± SD. AOH, acute ocular hypertension