Journal: PLoS ONE

Article Title: Novel Nongenomic Signaling by Glucocorticoid May Involve Changes to Liver Membrane Order in Rainbow Trout

doi: 10.1371/journal.pone.0046859

Figure Lengend Snippet: Rainbow trout hepatocytes were incubated either with cortisol (0, 100 or 1000 ng/mL) or benzyl alcohol (BA; 25 mM) for 10 min. Cell homogenates (40 µg protein) were probed with polyclonal rabbit antibody (Cell Signaling Technology, Beverly, MA) to either phospho-(Ser) PKC substrate (A), phospho-PKA Substrate (RRXS/T) (B) or phospho-Akt substrate (RXXS/T) (C). Equal loading was confirmed with β-actin (monoclonal mouse antibody; Sigma, St. Louis, MO). A representative immunoblot for each is shown; values are plotted as % control and shown as mean ± S.E.M (n = 3 independent fish); bars with different letters are significantly different (repeated measures ANOVA, p<0.05). *significantly different from control (Paired Student’s t-test; p<0.05).

Article Snippet: This was followed with an overnight incubation (1∶1000 dilution) with either phospho-(Ser) PKC substrate, phospho-Akt substrate (RXXS/T) or phospho-PKA Substrate (RRXS/T) polyclonal rabbit antibodies (Cell Signaling Technology, Beverly, MA).

Techniques: Incubation, Western Blot