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ACCEPTED MANUSCRIPT two NADPH oxidases, Nox1 and Nox4. Nox1 gene expression significantly increased in activated Müller cells compared to controls, while the expression of Nox4 did not change significantly (Figure 3B, C). Treatment with 670nm light significantly reduced the expression of both Nox1 and Nox4 in activated Müller cells (P<0.05, Figure 3B, C). Another hallmark of neurodegenerative diseases is the activation of the immune response (Chen and Xu, 2012). We examined the expression of CCL2, a potent chemokine produced by Müller cells shortly after retinal injury, as well as IL-6, a pro-inflammatory cytokine. Müller cells activated by the supernatant of PD-damaged 661W cells showed an increased gene and protein expression of CCL2 (Figure 3D, F). Treatment with 670nm light reduced CCL2 expression significantly (Figure 3D, F), confirming our in vivo findings (Figure 1B, G). Il-6 gene expression did not change in activated Müller cells, but was significantly reduced by 670nm irradiation (P<0.05, Figure 3E). IL-6 protein levels were significantly lower in activated Müller cells than controls, and 670nm treatment reduced IL-6 protein levels further (P<0.05, Figure 3G). 3.4 670nm light treatment has no effect on NLRP3 inflammasome activation in Müller cells NLRP3 inflammasome activation has been found in retinal degenerations including AMD (Doyle et al., 2012a; Tarallo et al., 2012). Using the in vivo PD model, we have previously shown an increase in the expression of IL-1β in recruited microglia and macrophages and subsequent NLRP3 inflammasome activation (Natoli et al., 2017b). Using our in vitro model, we investigated whether this inflammasome activation is Müller cell related, and if 670nm light treatment can influence inflammasome activation by its direct effect on Müller cells. The gene expression levels of Casp8, Nlrp3, Pycard, Casp1 and Il-18 were not significantly different in the activated Müller cells compared to controls. Treatment 14 ACCEPTED MANUSCRIPTPDF Image | Photobiomodulation with 670 nm light ameliorates retinal
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