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. Author manuscript; available in PMC: 2013 Dec 1.
Published in final edited form as: Int J Biochem Cell Biol. 2012 Aug 22;44(12):2106–2115. doi: 10.1016/j.biocel.2012.08.016

Figure 5.

Figure 5

Replenishment of intracellular GSH reverses the inhibitory effect of GSH depletion on LIF-induced STAT3 and JAK1 activation. Cardiac myocytes were treated for 6 h with different concentrations of BSO (lanes 3,4,7,8), washed 2× with medium, and incubated for 24 h in serum-free medium containing vehicle or 2 mM glutathione monoethyl ester (GME) (lanes 5-8). Cells were dosed with vehicle (lanes 1 & 5) or 2 ng/mL LIF (lanes 2-4 & 6-8) for 10 min. (A) Cell lysates were prepared and analyzed by Western immunoblotting for STAT3 Y705 phosphorylation. Membranes were stripped and reprobed for total STAT3 and GAPDH to confirm equal loading. (B) Immunoprecipitates of JAK1 from cell lysates were immunoblotted for tyrosine phosphorylated JAK1 (upper) and total JAK1 (lower) to ensure equal loading. Blots shown are representative of 5 independent experiments. (C) Densitometric analysis of immunoreactive bands. *P < 0.05 and ***P < 0.001 vs. maximal activation with LIF, ANOVA and Dunnett’s multiple comparison test.