Figure 3.

Knockdown of PBRM1 Results in Elevated ROS under Cellular Stress Conditions

(A) NMuMG cells were trypsinized and stained for 30 min with H₂-DCFDA, washed with PBS, and analyzed using flow cytometry. The mean fluorescence value for 10,000 cells was calculated from four independent experiments.

(B) NMuMG cells grown in 96-well plates were treated with increasing concentrations of H₂O₂ for 1 h, washed with PBS, and incubated with H₂-DCFDA for 30 min. Reagent was washed away, and the DCF fluorescence was measured in live cells.

(C) NMuMG cells grown in 96-well plates were treated with media containing varying concentrations of glucose (normal media = 25 mM) for 16 h, washed with PBS, and incubated with H₂-DCFDA for 30 min. Reagent was washed away, and the DCF fluorescence was measured in live cells.

(D) NMuMG cells grown in 96-well plates were treated with media containing varying concentrations of CoCl₂ for 24 h, washed with PBS, and incubated with H₂-DCFDA for 30 min. Reagent was washed away, and the DCF fluorescence was measured in live cells.

(E) NMuMG cells grown in 96-well plates were treated with media containing varying concentrations of doxorubicin for 24 h, washed with PBS, and incubated with H₂-DCFDA for 30 min. Reagent was washed away, and the DCF fluorescence was measured in live cells.

*p < 0.05, **p < 0.01, ***p < 0.001 (paired Student's t test). ns, not significant. Error bars represent S.D.