factor 2  (Bioss)

Journal: Antioxidants

Article Title: Nrf2 Activated by PD-MSCs Attenuates Oxidative Stress in a Hydrogen Peroxide-Injured Retinal Pigment Epithelial Cell Line

doi: 10.3390/antiox14111279

Figure Lengend Snippet: PD-MSC cocultivation activates the Nrf2 signaling pathway in ARPE-19 cells exposed to H 2 O 2 . mRNA expression of ( A ) PI3K p110a, ( B ) AKT and ( C ) Nrf2 levels in H 2 O 2 -treated ARPE-19 cells treated with 10 ng/mL PEDF, cocultured PD-MSCs or their combination. ( D ) Western blot analysis of antioxidant enzymes in cell lysates of H 2 O 2 -treated ARPE-19 cells cocultured with PD-MSCs. Quantification of ( E ) PI3K p110a, ( F ) AKT, ( G ) Nrf2 and ( H ) KEAP1 in cell lysates of H 2 O 2 -treated ARPE-19 cells cocultured with PD-MSCs. Quantitative analysis and representative images of ( I , J ) phospho-Nrf2 and ( K , L ) KEAP1 in H 2 O 2 -treated ARPE-19 cells cocultured with 10 ng/mL PEDF, PD-MSCs or their combination. The negative correlation between phospho-Nrf2 and KEAP1 was determined by the intensity of IF staining. DAPI was used for counterstaining. Scale bar = 50 µm. The data represent 2–3 independent experiments for each group and are expressed as the mean ± SEM. Statistical significance was determined by one-way ANOVA (* p < 0.05 vs. control, # p < 0.05 vs. H 2 O 2 treatment, $ p < 0.05 vs. PD-MSCs cocultivation).

Article Snippet: The following primary antibodies were used: anti-RDH11 (1:1000, bs-6214R, Bioss, Woburn, MA, USA), anti-RPE65 (1:1000, MA1-16578, InvitrogenTM, Carlsbad, CA, USA), anti-HMOX1 (1:1000, NBP1-97507, Novus Biologicals, Centennial, CO, USA), anti-SOD1 (1:1000, 4266S, Cell Signaling Technology, Danvers, MA, USA), anti-catalase (1:1000, ab52477, Abcam, Cambridge, UK), anti-phospho DRP1 (1:1000, PA5-64821, InvitrogenTM, Carlsbad, CA, USA), anti-OPA1 (ab157457, Abcam, Cambridge, UK), anti-ABCA1 (1:1000, NB400-105, NOVUS, Centennial, CO, USA), anti-ApoE (1:1000, A0304, ABclonal, Woburn, MA, USA), anti-PI3K p110α (1:1000, 4255s, Cell Signaling Technology, Danvers, MA, USA), anti-phospho AKT (1:1000, 9271S, Cell Signaling Technology, Danvers, MA, USA), anti-total AKT (1:1000, 9272S, Cell Signaling Technology, Danvers, MA, USA), anti-phospho Nrf2 (1:1000, BS2013R, Bioss, Woburn, MA, USA), anti-total Nrf2 (1:1000, BS1074R, Bioss, Woburn, MA, USA), anti-KEAP1 (1:1000, 8047S, Cell Signaling Technology, Danvers, MA, USA) and anti-GAPDH (1:1000, LF-PA0018, AbFrontier, Seoul, Republic of Korea).

Techniques: Expressing, Western Blot, Staining, Control

Journal: Antioxidants

Article Title: Nrf2 Activated by PD-MSCs Attenuates Oxidative Stress in a Hydrogen Peroxide-Injured Retinal Pigment Epithelial Cell Line

doi: 10.3390/antiox14111279

Figure Lengend Snippet: Summary illustration of the therapeutic effects of PD-MSCs on H 2 O 2 -injured RPE by increasing antioxidant enzymes via the Nrf2 pathway. PD-MSCs activate Nrf2 through the PI3K/AKT signaling pathway, promoting the expression of antioxidant enzymes. Mitochondrial dynamics are regulated, leading to reduced mitochondrial ROS levels and stabilized mitochondrial membrane potential. RPE regeneration is enhanced with decreased lipoprotein accumulation and reduced apoptosis.

Article Snippet: The following primary antibodies were used: anti-RDH11 (1:1000, bs-6214R, Bioss, Woburn, MA, USA), anti-RPE65 (1:1000, MA1-16578, InvitrogenTM, Carlsbad, CA, USA), anti-HMOX1 (1:1000, NBP1-97507, Novus Biologicals, Centennial, CO, USA), anti-SOD1 (1:1000, 4266S, Cell Signaling Technology, Danvers, MA, USA), anti-catalase (1:1000, ab52477, Abcam, Cambridge, UK), anti-phospho DRP1 (1:1000, PA5-64821, InvitrogenTM, Carlsbad, CA, USA), anti-OPA1 (ab157457, Abcam, Cambridge, UK), anti-ABCA1 (1:1000, NB400-105, NOVUS, Centennial, CO, USA), anti-ApoE (1:1000, A0304, ABclonal, Woburn, MA, USA), anti-PI3K p110α (1:1000, 4255s, Cell Signaling Technology, Danvers, MA, USA), anti-phospho AKT (1:1000, 9271S, Cell Signaling Technology, Danvers, MA, USA), anti-total AKT (1:1000, 9272S, Cell Signaling Technology, Danvers, MA, USA), anti-phospho Nrf2 (1:1000, BS2013R, Bioss, Woburn, MA, USA), anti-total Nrf2 (1:1000, BS1074R, Bioss, Woburn, MA, USA), anti-KEAP1 (1:1000, 8047S, Cell Signaling Technology, Danvers, MA, USA) and anti-GAPDH (1:1000, LF-PA0018, AbFrontier, Seoul, Republic of Korea).

Techniques: Expressing, Membrane