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. Author manuscript; available in PMC: 2020 Aug 1.

Published in final edited form as: Free Radic Biol Med. 2019 May 16;139:70–79. doi: 10.1016/j.freeradbiomed.2019.05.019

Fig 6. — Alternol increases XDH protein expression and induces cellular protein damages. A; Exponentially grown cells from each line as indicated were harvested for whole cell lysis. Equal amount of proteins was used in anti-XDH western blot assay. Actin blot served as protein loading control.

B-D; After treatment with Alternol or the solvent as indicated or at 10 μM for 4h, cells were harvested in cold PBS and cellular proteins were subjected to anti-XDH western blot assay. Actin blot served as protein loading control.

E-G; After treatment with Alternol (10 μM), N-Ac (5 μM) or Febuxostat (20 μM) as indicated, cells were harvested in cold PBS and cellular proteins were extracted in G-lysis buffer as described in the text. Biotin derivatized proteins were separated on 15% SDS-PAGE gel and visualized using HRP-linked Biotin antibodies.

H. After Alternol treatment as indicated, cells were harvested in cold PBS and subjected to 3-NT labeling and measurement as described [28]. Quantitative data were summarized from three independent measurements. The asterisk indicates a statistical significance compared to the DMSO control (Student t-test, p <0.01).

Fig 6. — Alternol increases XDH protein expression and induces cellular protein damages. A; Exponentially grown cells from each line as indicated were harvested for whole cell lysis. Equal amount of proteins was used in anti-XDH western blot assay. Actin blot served as protein loading control.

B-D; After treatment with Alternol or the solvent as indicated or at 10 μM for 4h, cells were harvested in cold PBS and cellular proteins were subjected to anti-XDH western blot assay. Actin blot served as protein loading control.

E-G; After treatment with Alternol (10 μM), N-Ac (5 μM) or Febuxostat (20 μM) as indicated, cells were harvested in cold PBS and cellular proteins were extracted in G-lysis buffer as described in the text. Biotin derivatized proteins were separated on 15% SDS-PAGE gel and visualized using HRP-linked Biotin antibodies.

H. After Alternol treatment as indicated, cells were harvested in cold PBS and subjected to 3-NT labeling and measurement as described [28]. Quantitative data were summarized from three independent measurements. The asterisk indicates a statistical significance compared to the DMSO control (Student t-test, p <0.01).