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. Author manuscript; available in PMC: 2015 Mar 1.
Published in final edited form as: Free Radic Biol Med. 2013 Dec 1;68:148–158. doi: 10.1016/j.freeradbiomed.2013.11.018

Figure 5. Effects of Ubc9 overexpression on LPS-mediated suppression of Gclc and Gclm expression, and sumoylation of MafG and Nrf2 in RAW cells.

Figure 5

Part A) shows the effect of Ubc9 overexpression on basal promoter activity and after LPS treatment. Ubc9 overexpression raised basal Gclc (but not Gclm) promoter activity and prevented LPS-mediated inhibition of both promoter activity. Results are from three experiments, expressed as % of respective basal promoter activity, *p<0.05 vs. Gclc basal promoter activity, †p<0.05 vs. Gclc+LPS, **p<0.05 vs. Gclm basal promoter activity. Part B) shows the effect of overexpressing Ubc9 or empty vector (V) on Ubc9, Gclc and Gclm mRNA levels at baseline and after LPS treatment. Results are from three experiments, *p<0.05 vs. V, †p<0.05 vs. LPS. Part C) shows the same experimental design as part B on protein levels of the same genes. Representative Western blots are shown with densitometric values shown below the blots expressed as % of V. Note the higher molecular weight band of Ubc9-HA. Both Ubc9 bands were included in their densitometric measurement and all were normalized to actin. Part D) shows nuclear levels of total and SUMO-1 sumolyated MafG and Nrf2. Results are mean±SE from three experiments expressed as % of V, *p<0.05 vs. V, †p<0.02 vs. LPS.