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ACCEPTED MANUSCRIPT did not alter ∆Ψm in control cells, but increased in activated Müller cells, compared to untreated activated cells (P<0.05, Figure 5E). 3.6 670nm light regulated Müller cell-mediated activation of microglia and macrophages (MG/MΦ) following PD After incubation with control supernatant (SN) (Figure 6B), or 670nm SN (Figure 6C), primary MG/MΦ displayed a ramified morphology with defined processes, indicating a resting state. However, the shape of MG/MΦ became more amoeboid with smaller processes following incubation with SN from activated Müller cells (PD SN, Figure 6D), indicating activation of these MG/MΦ. After incubation with SN from 670-treated activated Müller cells (PD+670nm SN, Figure 6E), MG/MΦ were more ramified and processes extending from cell soma were apparent, indicating that 670nm reduced activation of MG/MΦ. We used flow cytometry to detect the fluorescence intensity of CD11b (Figure 6F-G) and CD86 expression (Figure 6H-I) after stimulation with differing SN to assess their activation state. MG/MΦ had a higher expression of CD11b and CD86 after incubation with PD SN compared to cells incubated with PD+670nm SN (P<0.05, Figure 6F-I), suggesting that PBM reduced macrophage activation. To further confirm MG/MΦ activation, we assessed the gene expression of Ccl2, Il-1β, Tnfα, Il-6, Il-10 and Sod2, which are all markers of macrophage activation (Figure 6J-O). The expression of these genes was increased in MG/MΦ incubated in PD SN, indicating that supernatant from PD-stressed Müller cells have an activating effect on these cells. Treatment with 670nm significantly lowered MG/MΦ activation as suggested by the reduced expression of these genes (P<0.05, Figure 6J-O). 4. Discussion 16 ACCEPTED MANUSCRIPTPDF Image | Photobiomodulation with 670 nm light ameliorates retinal
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