m akt inhibitor viii  (Santa Cruz Biotechnology)

Journal: The Journal of Biological Chemistry

Article Title: Pleckstrin Homology (PH) Domain Leucine-rich Repeat Protein Phosphatase Controls Cell Polarity by Negatively Regulating the Activity of Atypical Protein Kinase C *

doi: 10.1074/jbc.M116.740639

Figure Lengend Snippet: Inhibition of Akt does not affect cell polarity. A, stable sh-Con, sh-PHLPP1, and sh-PHLPP2 Caco2 cells seeded in 3D matrix as a single cell suspension. At day 7, the cells were left untreated or treated with Akt-VIII (0.5 μm) or PKCζ inhibitor (PKCζ-i, 20 μm) for an additional 7 days. The cells were subsequently fixed and stained with Alexa Fluor 488-conjugated phalloidin (green) and DAPI (blue). Scale bars, 10 μm. B, the sizes of 50 randomly chosen cysts as shown in A were measured using the Nikon Elements AR software. The size distribution is shown in the box-whisker plot. The average cyst sizes for the following cells are (means ± S.E., in μm): sh-Con, 84.2 ± 4.7; sh-Con+Akt-VIII, 59.6 ± 4.0; sh-Con+PKCζ-i, 61.7 ± 4.8; sh-PHLPP1, 122.1 ± 7.2; sh-PHLPP1+Akt-VIII, 55.7 ± 2.6; sh-PHLPP1+PKCζ-i, 59.1 ± 3.2; sh-PHLPP2, 108.2 ± 6.7; sh-PHLPP2+Akt-VIII, 59.3 ± 3.2; and sh-PHLPP1+PKCζ-i, 73.1 ± 6.5 (# indicates p < 0.001 and * indicates p < 0.01 when compared with untreated sh-Con cells, ¶ indicates p < 0.001 when compared with untreated sh-PHLPP1 cells, and § indicates p < 0.001 when compared with untreated sh-PHLPP2 cells by Student's t test). C, the percentages of cysts with a single lumen were quantified based on the pattern of actin staining and expressed graphically. Fifty randomly chosen cysts from each group were scored. The shaded bars represent cysts with one lumen, and the open bars represent cysts with multiple or filled lumens.

Article Snippet: Akt inhibitor VIII (0.5 μ m ), myristoylated-PKCζ pseudosubstrate inhibitor (20 μ m ), and PI3K inhibitor {"type":"entrez-nucleotide","attrs":{"text":"LY294002","term_id":"1257998346","term_text":"LY294002"}} LY294002 (20 μ m ) were purchased from Millipore.

Techniques: Inhibition, Staining, Software, Whisker Assay

Journal: International Journal of Endocrinology

Article Title: Effect of Gelam Honey on the Oxidative Stress-Induced Signaling Pathways in Pancreatic Hamster Cells

doi: 10.1155/2013/367312

Figure Lengend Snippet: Effect of quercetin and Gelam honey extract on pAkt (Ser473) expression.Quantitative analysis and representative western blot analysis of pAkt (Ser473) in HIT-T15 cells pretreated with quercetin and honey extract in cells cultured in 20 mM ((a), (c), (e)) and 50 mM ((b), (d), (f)) glucose. A sustained increase in the level of pAkt (ser473) was observed after pretreatment with quercetin and honey extract. Akt inhibitor VIII prevented the expression Akt ser473 phosphorylation induced by quercetin and honey extract. The results were normalized with β actin antibody. Data were presented as the mean ± standard deviation. (e) * P < 0.05; # P < 0.005 quercetin and honey extract treated compared to the 20 mM glucose alone. (f) * P < 0.05; # P < 0.005 quercetin and honey extract treated compared to the 50 mM glucose alone.

Article Snippet: To investigate inhibitory effects on Akt signaling pathway, cells were incubated with 5 μ M Akt inhibitor VIII (Santa Cruz Biotechnology sc-203173) for one hour before pretreatment with quercetin and honey extract.

Techniques: Expressing, Western Blot, Cell Culture, Phospho-proteomics, Standard Deviation

Journal: International Journal of Endocrinology

Article Title: Effect of Gelam Honey on the Oxidative Stress-Induced Signaling Pathways in Pancreatic Hamster Cells

doi: 10.1155/2013/367312

Figure Lengend Snippet: The effect of flavonoids and Gelam honey extract on insulin content. (a) Effect of pretreatment with quercetin and Gelam honey extract and the addition of Akt inhibitor VIII on the insulin content in cells cultured in 20 mM glucose. There was a significant increase in insulin content (* P < 0.05) when the cells were pretreated with quercetin and honey. There was a significant decrease in insulin content (* P < 0.05) when the cells were treated with Akt inhibitor VIII, before pretreating with quercetin and Gelam honey extract. (b) Effect of pretreatment with quercetin and Gelam honey and the addition of Akt inhibitor VIII on the insulin content in cells cultured in 50 mM glucose. There was a significant increase in insulin content (* P < 0.05, # P < 0.005) when the cells were pretreated with quercetin and honey. There was a significant decrease in insulin content (* P < 0.05, # P < 0.005) when the cells were treated with Akt inhibitor VIII, before pretreating with quercetin and Gelam honey extract.

Article Snippet: To investigate inhibitory effects on Akt signaling pathway, cells were incubated with 5 μ M Akt inhibitor VIII (Santa Cruz Biotechnology sc-203173) for one hour before pretreatment with quercetin and honey extract.

Techniques: Cell Culture