Figure 5
The effect of LDM DOX administration and SN co-supplementation on molecular markers of oxidative stress and the anti-oxidant response in skeletal muscle. Western blotting experiments were undertaken utilizing soleus muscle homogenate. When probing for molecular markers of oxidative stress, there was no significant change in (A) 4-HNE protein expression, a marker of lipid peroxidation, between treatment groups, but there was a significant increase in (B) nitrotyrosine protein expression, a marker of excessive peroxynitrite (ONOO-) production in DOX and DOX + SN treated mice (*p < 0.05) compared to VEH. There was also a significant increase in (C) NRF-2 protein expression, the master regulator of the anti-oxidant response to cellular stress, in DOX and DOX + SN treated mice (*p < 0.05) compared to VEH. (D) DJ-1 protein expression was probed for as a positive regulator of NRF-2 activation, which was interestingly, shown to increase in DOX mice (*p < 0.05) compared to VEH, whilst there was no significant change in DOX + SN treated mice. Negative regulators of NRF-2, i.e. (E) Keap-1, (F) phosphorylated p62ser349 and total p62 (depicted as a ratio of the total), were not significantly changed from treatment. Downstream targets of the anti-oxidant response, i.e. (G) NQO-1, (H) SOD-1 and (I) HO-1 did not significantly change from treatment. (J) Representative images of the antibodies are displayed alongside a representative image of Coomassie Blue used to normalize to total protein content. n = 5–8.
