Journal: Investigative Ophthalmology & Visual Science

Article Title: TFEB-Mediated Lysosomal Restoration Alleviates High Glucose-Induced Cataracts Via Attenuating Oxidative Stress

doi: 10.1167/iovs.63.6.26

Figure Lengend Snippet: Curcumin analog C1 exerted protective effects in HG-induced cataracts. Rat lenses were subjected to control (5.5 mM), HG (50 mM), and HG (50 mM) with C1 (5 µM) medium for 7 days. ( A ) Representative images of cultured rat lenses and the percentage distribution of cataract scores in the control, HG, and HG + C1 groups. ( B ) TEM showed that C1 treatment alleviated abnormally large autophagic vesicles ( arrows ) with massive undegraded substrates in anterior capsular LECs under HG conditions. ( C ) Protein expression levels and quantitative analysis of α-SMA, FN, LC3-I/II, and P62 determined by Western blotting in LECs. ( D ) Protein expression levels and quantitative analysis of LC3-I/II and P62 determined by Western blotting in LFCs. All the data were shown as mean ± standard deviation. * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: To study the effects of TFEB activation, we treated lenses with curcumin analog C1 (C1; 5 μM; Selleck) or tetrahydrocurcumin (THC; 5 μM; Selleck) upon HG (50 mM) treatment.

Techniques: Control, Cell Culture, Expressing, Western Blot, Standard Deviation

Journal: Investigative Ophthalmology & Visual Science

Article Title: TFEB-Mediated Lysosomal Restoration Alleviates High Glucose-Induced Cataracts Via Attenuating Oxidative Stress

doi: 10.1167/iovs.63.6.26

Figure Lengend Snippet: C1 enhanced lysosomal degradation via promoting the nuclear translocation of TFEB in HG-cultured LECs. Primary rabbit LECs were exposed to control (5.5 mM), HG (25 mM), and HG (25 mM) with C1 (5 µM) medium for 3 days. ( A ) ROS levels were detected by DCFH-DA assay. ( B , C ) Protein expression levels and quantitative analysis of α-SMA, FN, LC3-I/II, and P62 determined by Western blotting in LECs. ( D ) Representative fluorescence images of mCherry-EGFP-LC3 puncta within LECs and quantification. ( E ) Immunofluorescence staining and Western blot analysis ( F ) showed that C1 increased the expression levels and distributions of TFEB in the cytoplasm and nuclei in LECs. ( G ) Protein expression levels and quantitative analysis of Pro-CTSB, LAMP1, and ubiquitin determined by Western blotting in LECs. ( H ) Lysotracker staining and immunofluorescence staining of LAMP1 puncta in LECs. Relative protein levels of total proteins were standardized to the expression of β-Actin, whereas nuclear proteins were normalized to Lamin B1. All the data were shown as mean ± standard deviation. * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: To study the effects of TFEB activation, we treated lenses with curcumin analog C1 (C1; 5 μM; Selleck) or tetrahydrocurcumin (THC; 5 μM; Selleck) upon HG (50 mM) treatment.

Techniques: Translocation Assay, Cell Culture, Control, DCFH-DA Assay, Expressing, Western Blot, Fluorescence, Immunofluorescence, Staining, Ubiquitin Proteomics, Standard Deviation

Journal: Investigative Ophthalmology & Visual Science

Article Title: TFEB-Mediated Lysosomal Restoration Alleviates High Glucose-Induced Cataracts Via Attenuating Oxidative Stress

doi: 10.1167/iovs.63.6.26

Figure Lengend Snippet: Comparison of THC and C1 in alleviating HG-induced cataracts. Primary rabbit LECs were exposed to control (5.5 mM), HG (25 mM), HG (25 mM) with THC (5 µM), and HG (25 mM) with C1 (5 µM) medium for 3 days. ( A ) Protein expression levels and quantitative analysis of TFEB determined by Western blotting in LECs. ( B ) DCFH-DA assay compared ROS levels in the control, HG, HG + THC, and HG + C1 groups. Rat lenses were subjected to control (5.5 mM), HG (50 mM), HG (50 mM) with THC (5 µM), and HG (50 mM) with C1 (5 µM) medium for 7 days. ( C ) Representative images of cultured rat lenses and the percentage distribution of cataract scores in the control, HG, HG + THC, and HG + C1 groups. ( D ) Protein expression levels and quantitative analysis of α-SMA, FN, LC3-I/II, and P62 determined by Western blotting in LECs. All the data were shown as mean ± standard deviation. * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: To study the effects of TFEB activation, we treated lenses with curcumin analog C1 (C1; 5 μM; Selleck) or tetrahydrocurcumin (THC; 5 μM; Selleck) upon HG (50 mM) treatment.

Techniques: Comparison, Control, Expressing, Western Blot, DCFH-DA Assay, Cell Culture, Standard Deviation