Journal: Mediators of Inflammation

Article Title: Kaempferide Protects against Myocardial Ischemia/Reperfusion Injury through Activation of the PI3K/Akt/GSK-3 β Pathway

doi: 10.1155/2017/5278218

Figure Lengend Snippet: Kae treatment increased the levels of phosphorylated Akt and phosphorylated GSK-3 β in the myocardium. PI3K inhibition (LY294002) attenuates Kae-induced increases in phospho-Akt and phospho-GSK-3 β levels. (a) Representative Western blots for Akt, p-Akt, GSK-3 β , and P-GSK-3 β . (b) Grey value analysis demonstrates that Kae treatment increases the ratio of p-Akt/Akt. (c) Grey value analysis demonstrates that Kae treatment increases the ratio of p-GSK-3 β /GSK-3 β . ## P < 0.01 versus sham; ∗∗ P < 0.01 versus I/R; ++ P < 0.01 versus H-Kae + I/R.

Article Snippet: Membranes were then incubated first with the following primary antibodies: rabbit anti-rat GSK-3 β , rabbit anti-rat P-GSK-3 β , rabbit anti-rat Akt, rabbit anti-rat p-Akt, rabbit anti-rat Nrf2, rabbit anti-rat cleaved caspase-3, rabbit anti-rat caspase-3, and rabbit anti-rat β -actin polyclonal antibodies (Cell Signaling, Beverly, MA, USA) at 4°C overnight.

Techniques: Inhibition, Western Blot

Journal: International Journal of Endocrinology

Article Title: Ghrelin Increases Beta-Catenin Level through Protein Kinase A Activation and Regulates OPG Expression in Rat Primary Osteoblasts

doi: 10.1155/2015/547473

Figure Lengend Snippet: Representative Western blot (a) and quantification (b) of timecourse experiments performed on rat osteoblast-like cells treated with ghrelin (AG, 10 −10 M) for 15, 30, 60, and 90 min. Phosphorylated-GSK-3 β (P-GSK-3 β ) levels were measured in whole cell lysates and normalized to total GSK-3 β levels (Tot-GSK3 β ). Results are shown as ratio of P-GSK3 β measured in treated versus untreated cells. Data are the mean ± SEM of 5 experiments * P < 0.05, versus untreated cells.

Article Snippet: Western blots were performed using a specific antibody against rat β -catenin diluted 1 : 5000 in 5% milk in Tris–HCl with 0.1% Tween-20 (TBST); rat P-GSK3 β (1 : 2000 5% milk in TBST); rat total GSK3 β (1 : 2000 5% milk in TBST) (Cell Signaling Technology, Boston, MA) or against rat β -actin (1 : 2000 5% milk in TBST) (Santa Cruz Biotechnology, Inc., Heidelberg, Germany).

Techniques: Western Blot

Journal: International Journal of Endocrinology

Article Title: Ghrelin Increases Beta-Catenin Level through Protein Kinase A Activation and Regulates OPG Expression in Rat Primary Osteoblasts

doi: 10.1155/2015/547473

Figure Lengend Snippet: Effect of the pretreatment with the PKA inhibitor H89 (2 × 10 −6 M, 30 min before) on (a) β -catenin and (b) phosphorylated-GSK3 β (P-GSK3 β ) levels measured in whole cell lysates by Western blot on rat osteoblast-like cells (rOB) treated with ghrelin (AG, 10 −10 M for 1 h). β -catenin levels were normalized to β -actin levels; P-GSK3 β was normalized to total GSK3 β levels (Tot-GSK3 β ). Results are expressed as ratio of β -catenin or P-GSK3 β measured in treated cells versus untreated cells. Insets : typical gels obtained from rOB cell lysates after treatment with ghrelin alone or together with H89. Data are the mean ± SEM of 5 experiments * P < 0.05 versus untreated cells; § P < 0.05 versus AG treated cells.

Article Snippet: Western blots were performed using a specific antibody against rat β -catenin diluted 1 : 5000 in 5% milk in Tris–HCl with 0.1% Tween-20 (TBST); rat P-GSK3 β (1 : 2000 5% milk in TBST); rat total GSK3 β (1 : 2000 5% milk in TBST) (Cell Signaling Technology, Boston, MA) or against rat β -actin (1 : 2000 5% milk in TBST) (Santa Cruz Biotechnology, Inc., Heidelberg, Germany).

Techniques: Western Blot