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. Author manuscript; available in PMC: 2009 Oct 26.
Published in final edited form as: Am J Physiol Endocrinol Metab. 2007 Oct 9;293(6):E1552–E1563. doi: 10.1152/ajpendo.00134.2007

Fig. 3. Total and pS293 phosphorylated BCKD E1α and BCKD kinase immunoreactivity in liver from ob/ob mice.

Fig. 3

Fig. 3

Fig. 3

Fig. 3

Liver protein lysates, made from frozen lean (L) and ob/ob (O) rat liver powders, were equalized for protein content, separated by SDS-PAGE and transferred to PVDF for immunoblotting. Panel A shows representative immunoblots of total BCKD E1α (liver does not express BCATm), pS293 E1α, BCKD kinase and β-Actin (loading control). Other panels show densitometric analysis of replicates for BCKD E1α (Panel B), pS293 E1α/BCKD E1α ratio (Panel C) and BCKD kinase (Panel D). Fed and Fasted data were initially derived from two separate blots at different times. To compare them together, the remaining samples were electrophoresed on the same blot and reanalyzed together, however there was insufficient material to rerun the BCKD kinase. The original data are shown for that endpoint; a t-test was used to compare the lean and obese states only (*** p < 0.001, n=6/group). The other reanalyzed samples were compared between lean and obese as well as nutritional status using an ANOVA. Asterisks indicate where statistical differences between the lean and obese condition in the corresponding nutritional state was observed based upon a Student-Newman-Keuls Multiple Comparisons post-test (*P<0.05, *** p < 0.001). No differences were observed between the lean and fasted states for any antigen. Western blotting for β-Actin was used as a loading control. The densitometry values were 7539 ± 665; 7358 ± 692; 6835 ± 532 and 6833 ± 578 for fed lean, fed obese, fasted lean and fasted obese mouse liver, respectively (N.S., p>0.05).