Journal: Molecules

Article Title: The Effects of 2′,4′-Dihydroxy-6′-methoxy-3′,5′- dimethylchalcone from Cleistocalyx operculatus Buds on Human Pancreatic Cancer Cell Lines

doi: 10.3390/molecules24142538

Figure Lengend Snippet: Effect of DMC on caspase activation and PARP degradation protein expression in PANC-1 cells. Cells were treated with DMC (3–30 µM) for 48 h. Protein 50 µg/lane from cells lysates were electrophoresed on SDS-PAGE gels, then transferred to total blot PVDF membranes. β-Actin was used as a control, (–), 0.1% DMSO-treated cells. The experiments were carried out in three replicates.

Article Snippet: Protein contents were normalized by reprobing the same membrane with anti-β-actin detection; previously used membranes were soaked in stripping buffer (Gene Bio-Application Ltd., Yavne, Israel) at room temperature for 20 min.

Techniques: Activation Assay, Expressing, SDS Page

Journal: Molecules

Article Title: The Effects of 2′,4′-Dihydroxy-6′-methoxy-3′,5′- dimethylchalcone from Cleistocalyx operculatus Buds on Human Pancreatic Cancer Cell Lines

doi: 10.3390/molecules24142538

Figure Lengend Snippet: Effect of DMC on Bcl-2, Bax and Cyt-c protein expression in PANC-1 cells. Cells were treated with DMC (3–30 µM) for 48 h. Protein 50 µg/lane from cells lysates were electrophoresed on SDS-PAGE gels, then transferred to total blot PVDF membranes. β-Actin was used as a control, (–) 0.1% DMSO-treated cells. The experiments were carried out in three replicates. * P < 0.05 and P < 0.01 compared with control group.

Article Snippet: Protein contents were normalized by reprobing the same membrane with anti-β-actin detection; previously used membranes were soaked in stripping buffer (Gene Bio-Application Ltd., Yavne, Israel) at room temperature for 20 min.

Techniques: Expressing, SDS Page

Journal: JID Innovations

Article Title: Supply Chain Disruptions During COVID Pandemic Uncover Differences in Keratinocyte Culture Media

doi: 10.1016/j.xjidi.2022.100151

Figure Lengend Snippet: Markers of KC differentiation are significantly impacted by propagation media. Cells were cultured in KSFM, KGM2 or Defined media, grown to confluency, and switched to DMEM media containing calcium (1.8 mM) to initiate differentiation. Cell lysates were collected from undifferentiated KC 24 hours after reaching confluency (a) and collected from differentiated KC at Day (D) 1, 2 and 3 post differentiation (b-e) . Differentiation markers cytokeratin 10 (CK10), loricrin (LOR), and tight junction proteins claudin-1 (CLDN1) and occludin (OCLN) were detected by Western blot analysis. A representative blot for each protein is provided (f) . Expression was quantified by densitometry with normalization to β-actin expression. n = 3 experiments, graphs with less than 3 points indicates protein was not detected (ND) in experiments. Solid bars indicate undifferentiated KC in respective media, open bars indicate KC that have been switched from respective media to DMEM. Friedman test with Dunn’s post-hoc test (pairwise) of KSFM vs KGM2 or KSFM vs Defined. Significance: *p<0.05. Data presented as median (Q1, Q3).

Article Snippet: Samples were normalized to β-actin expression and protein content (Pierce® BCA Protein Assay Kit).

Techniques: Cell Culture, Western Blot, Expressing

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Pretreatment Donors after Circulatory Death with Simvastatin Alleviates Liver Ischemia Reperfusion Injury through a KLF2-Dependent Mechanism in Rat

doi: 10.1155/2017/3861914

Figure Lengend Snippet: Simvastatin pretreatment maintains the expression of KLF2 and its protective target genes (a). The protein expression of KLF2 (b), phosphorylation eNOS (c), Bcl-2/Bax (d), and β -actin were determined by Western blotting and the gray values were calculated ( n = 6 per group; ∗ P < 0.05 versus NP group, # P < 0.05 versus DCD-Con group). The mRNA expression of KLF2 (e), eNOS (f), TM (g), and HO-1 (h) were assessed by RT-PCR. The experiments were repeated three times and the data are shown as mean ± SD ( n = 6; ∗ P < 0.05 versus NP group, # P < 0.05 versus DCD-Con group).

Article Snippet: All bands were assayed for β -actin (1 : 1000, rabbit anti- β -actin antibody, Proteintech, Manchester, UK) content as standardization of sample loading.

Techniques: Expressing, Phospho-proteomics, Western Blot, Reverse Transcription Polymerase Chain Reaction

Journal: Oncotarget

Article Title: Phase II clinical and exploratory biomarker study of dacomitinib in recurrent and/or metastatic esophageal squamous cell carcinoma.

doi: 10.18632/oncotarget.6056

Figure Lengend Snippet: Figure 5: The anti-tumor efficacy of dacomitinib in ESCC cell lines with different EGFR expression. The viability of TE2, TE3, and HCE4 cell lines were measured by proliferation A. and clonogenic assay B.. The relative cell viability (%) and colonies represents the percent growth as compared to the control group (no treatment). C. Expression of EGFR was analyzed by quantitative real-time PCR. Expression of each EGFR mRNA is presented as relative to the mRNA expression of the internal control gene β-actin. D. The protein levels were measured by Western blot after 2 h treatment of dacomitinib.

Article Snippet: Protein content was resolved by SDS-PAGE, and was transferred to nitrocellulose membranes (Millipore, Temecula, CA). p-EGFR (Y1068), EGFR, p-AKT(S473), AKT, p-ERK (T202/Y204), and ERK secondary antibodies were purchased from Cell Signaling Technology (Beverly, MA, USA) and β-actin from Santa Cruz biotechnology (Santa Cruz, CA, USA).

Techniques: Expressing, Clonogenic Assay, Control, Real-time Polymerase Chain Reaction, Western Blot