Research Paper Volume 10, Issue 10 pp 2954—2972

Melatonin prevents senescence of canine adipose-derived mesenchymal stem cells through activating NRF2 and inhibiting ER stress

Figure 5. Melatonin reduces ERS through activating NRF2 and ERAD and inhibiting NF-κB and SASP. (A) Relative transcript levels of ERAD-related genes in P11 control, melatonin-, luzindole+melatonin- and oltipraz-treated cADMSCs. (B) Western blot quantification of ERAD-related protein in control, melatonin-, luzindole+melatonin- and oltipraz-treated P11 cADMSCs. (C) Relative transcript levels of ERAD-related genes in negative control, mock, shRNA vector, melatonin-treated, shNrf2-1, and shNrf2-2 P3 cADMSCs. (D) Relative transcript levels of ERS-related genes in negative control, mock, vector, melatonin-treated mock, shNrf2-1, and shNrf2-2 P3 cADMSCs. (E) Western blot and multiple quantifications of ERS-related protein (NRF2) in the negative control, mock, vector, melatonin-treated mock, shNrf2-1, and shNrf2-2 P3 cADMSCs. (F) Relative transcript levels of ERS-related genes in control, melatonin-, MG-132+melatonin- and NMS-873+melatonin-treated P11 cADMSCs. (G) Western blot quantification of ERS-related genes in control, melatonin-, MG-132+melatonin- and NMS-873+melatonin-treated P11 cADMSCs. (H) Relative transcript levels of SASP-related genes (P50 and P65) in control, melatonin-, luzindole+melatonin- and oltipraz-treated cADMSCs. (I) Western blot quantification of SASP-related proteins (P65, p-P65, and IKK) in control, melatonin-, luzindole+melatonin- and oltipraz-treated P11 cADMSCs. (J) NF-κB activity in control, melatonin-, luzindole+melatonin- and oltipraz-treated P11 cADMSCs was detected using dual-luciferase assay (K) Relative transcript levels of SASP-related genes (P50 and P65) in negative control, mock, vector, melatonin-treated mock, shNrf2-1, and shNrf2-2 P3 cADMSCs.