Journal: Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine

Article Title: Overexpression of WDFY2 inhibits prostate cancer cell growth and migration via inactivation of Akt pathway.

doi: 10.1177/1010428317704821

Figure Lengend Snippet: Figure 4. WDFY2 influenced prostate cancer development via Akt pathway specifically. (a) and (b) Western blotting showed that overexpressing WDFY2 did not affect the expression of E-cadherin and SLUG. (c) Real-time RT-PCR analysis of the mRNA level of Snail, Slug, Twist, ZEB1, and ZEB2 in the DU145 cells transfected with control vector and WDFY2. (d) and (e) WDFY2 overexpression showed specific downregulation of pAkt (Ser473). The values were the mean ± SEM of three independent experiments, and the p value is shown from Student’s t-test analysis.

Article Snippet: Some manufacturers were responsible for the production of antibodies, including anti-WDFY2 antibody (SAB), anti-E-cadherin (610181; BD Transduction Laboratories), anti-phospho-GSK3β, anti-Akt, anti-phospho-Akt (Ser473), anti-STAT3, anti-pSTAT3 (Tyr705), anti-P65, anti-pP65 (Ser536), anti-b-actin (Sigma-Aldrich), and also anti-tubulin (Proteintech).

Techniques: Western Blot, Expressing, Quantitative RT-PCR, Transfection, Control, Plasmid Preparation, Over Expression

Journal: Shock (Augusta, Ga.)

Article Title: Restorative mechanisms regulating protein balance in skeletal muscle during recovery from sepsis

doi: 10.1097/SHK.0000000000000762

Figure Lengend Snippet: A-F: Representative Western blots and quantitation are presented for phosphorylated and/or total Akt, PRAS40, TSC2, AMPK, and REDD1. The final panel is a representative tubulin blot demonstrating equal protein loading. Loading controls (tubulin or actin) were run for all proteins examined, but only representative data are shown. Bar graphs: were quantified for phosphorylation of each protein normalized to the total amount of the respective protein, except for REDD1 where total protein was normalized to tubulin. Control values were set to 100 arbitrary units (AU). Values are mean ± SEM. *P < 0.05 sepsis-recovery compared to pair-fed controls values; n = 17 and 10, respectively.

Article Snippet: Antibodies included (Cell Signaling, Beverly, MA, unless otherwise noted): S6K1, S6K1 (Thr389), rpS6, rpS6 (Ser240/244 and Ser230/235), 4E-BP1 (Bethyl Laboratories, Montgomery, TX), 4E-BP1 (Ser65), eukaryotic elongation factor (eEF)2, eEF2 (Thr56), eEF2 kinase, eEF2K (Ser366; Dr. Chris Proud), ERK (extracellular signal-regulated kinses), ERK1/2 (Thr202/Tyr204), RSK (90 kDa ribosomal S6 kinase), RSK1/2 (Ser380), REDD1 (regulated in development and DNA damage responses; ProteinTech, Chicago, IL), Akt, Akt (Thr308 and Ser473), PRAS40 (proline-rich Akt substrate of 40 kDa), PRAS40 (Thr246), eIF4B, eIF4B (Ser422), AMP-activated protein kinase-α (AMPK), AMPKα (Thr172), tuberous sclerosis complex 2 (TSC2), TSC2 (Thr1462), Unc-51 like autophagy activating kinase 1 (ULK1), ULK1 (Ser757), p62 (aka SQSTM1), light-chain 3B (LC3B)-I and –II, and MyoD and myogenin.

Techniques: Western Blot, Quantitation Assay

Journal: Journal of Cancer

Article Title: Gambogic acid affects ESCC progression through regulation of PI3K/AKT/mTOR signal pathway

doi: 10.7150/jca.41115

Figure Lengend Snippet: Expression of apoptosis-related proteins and PI3K/AKT/mTOR pathway components in ESCC cells incubated with GA. (A) The expression levels of cleaved-PARP1, Bcl-2, Bax, cleaved-caspase 3 and cleaved-caspase9 were detected by using western blot analysis. (B) Western blotting was performed to analyze the levels of PI3K, p-AKT, p-mTOR, AKT, mTOR and PTEN.

Article Snippet: The following primary antibodies were applied in the experiments: cleaved-caspase3, cleaved-caspase9, cleaved-PARP1, phosphorylated AKT, Bcl-2, MMP2, MMP9 and β-actin (Proteintech Group, Wuhan, China); PARP1, BAX, PI3K, total AKT, phosphorylated AKT (ser473), p-mTOR (ser2448), cyclinB1 and cyclinD1 (Cell Signaling Technology, MA, USA). β-actin was chosen as a loading control in all experiments.

Techniques: Expressing, Incubation, Western Blot