Fig. 2: NAM reduces mitochondrial ROS levels by increasing antioxidant enzymes. | Experimental & Molecular Medicine

Fig. 2: NAM reduces mitochondrial ROS levels by increasing antioxidant enzymes.

From: Nicotinamide enhances osteoblast differentiation through activation of the mitochondrial antioxidant defense system

Fig. 2: NAM reduces mitochondrial ROS levels by increasing antioxidant enzymes.

a The oxidative stress in MC3T3-E1 cells was determined by CellROXTM reagent after 10 μM NAM treatment with osteogenic media for 7 d. b The fluorescence intensity of (a) was calculated via ImageJ. c Mitochondrial superoxide was detected by MitoSOXTM mitochondrial superoxide indicator in MC3T3-E1 cells after 10 μM NAM treatment in osteogenic media for 7 d. d The fluorescence intensity of (c) was measured by ImageJ. e–i The mRNA expression levels of Pgc1α and ROS scavenger enzymes in MC3T3-E1 cells cultured in osteogenic medium supplemented with or without 10 μM NAM for 4 and 7 d were determined by RT‒qPCR. j The mitochondrial fraction was isolated from the culture under the same conditions to measure SOD2 enzymatic activity. k–o To understand the influence of PGC1A on ROS scavenger enzyme gene expression, MC3T3-E1 cells transfected with siCtrl or siPgc1α were cultured in osteogenic medium with or without 10 μM NAM for 4 d. p MC3T3-E1 cells were cultured in osteogenic medium with or without 10 μM NAM for 7 d. The protein expression level of ROS scavenger enzymes was determined by immunoblot analysis. Data are expressed as mean ± SD. *P < 0.05. **P < 0.01. ***P < 0.001. ****P < 0.0001. ns, not significant.

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