miR-26a-5p Attenuates Oxidative Stress and Inflammation in Diabetic Retinopathy through the USP14/NF-κB Signaling Pathway

Figure 4

USP14 enhancement reverses the impact of miR-26a-5p upregulation on HG-induced Müller cell dysfunction. (a) Western blotting was performed to quantify USP14 protein expression in Müller cells of the NC mimics, miR-26a-5p mimics, miR-26a-5p mimics + pcDNA, and miR-26a-5p mimics + pcDNA/USP14 groups. (b–d) Measurement of SOD, CAT, and MDA levels in Müller cells of the NC mimics, miR-26a-5p mimics, miR-26a-5p mimics + pcDNA, and miR-26a-5p mimics + pcDNA/USP14 groups was performed using specific commercial kits. (e) CM-H2DCFDA staining was performed to measure ROS levels in Müller cells of the NC mimics, miR-26a-5p mimics, miR-26a-5p mimics + pcDNA, and miR-26a-5p mimics + pcDNA/USP14 groups. (f) Western blotting was performed to quantify cytochrome c protein levels in the cytoplasm and mitochondria of Müller cells of the NC mimics, miR-26a-5p mimics, miR-26a-5p mimics + pcDNA, and miR-26a-5p mimics + pcDNA/USP14 groups. (g) Western blotting for evaluating GFAP protein expression in Müller cells of the NC mimics, miR-26a-5p mimics, miR-26a-5p mimics + pcDNA, and miR-26a-5p mimics + pcDNA/USP14 groups. (h–k) ELISA was performed to measure (h) VEGF level and the accumulation of (i–k) proinflammatory cytokines (TNF-α, IL-1β, and IL-6) in Müller cells of the NC mimics, miR-26a-5p mimics, miR-26a-5p mimics + pcDNA, and miR-26a-5p mimics + pcDNA/USP14 groups. and .