Journal: Thoracic Cancer

Article Title: SERPINB5 is a prognostic biomarker and promotes proliferation, metastasis and epithelial‐mesenchymal transition ( EMT) in lung adenocarcinoma

doi: 10.1111/1759-7714.15013

Figure Lengend Snippet: SERPINB5 knockdown suppresses the epithelial‐mesenchymal transition (EMT) of lung adenocarcinoma (LUAD) cells (a) Immunofluorescence staining showed the changes in the expression of EMT‐associated genes E‐cadherin (green) and vimentin (red) in LUAD cells. Nuclei were counterstained with DAPI (blue). (b) Western blot assay of the expression of EMT markers in LUAD cells transfected with si‐SERPINB5.

Article Snippet: Anti‐SERPINB5 was purchased from Immunoway Company (catalog no.: YT2657, 1:1000), and anti‐SNAIL (ER1706‐22, 1:1000), anti‐N‐cadherin (ET1607‐37, 1:1000), anti‐vimentin (EM0401, 1:1000) and anti‐E‐cadherin (ET1607‐75, 1:1000) were purchased from Hangzhou HuaAn Biotechnology.

Techniques: Immunofluorescence, Staining, Expressing, Western Blot, Transfection

Journal: Signal transduction and targeted therapy

Article Title: Integrative multi-omics and drug-response characterization of patient-derived prostate cancer primary cells.

doi: 10.1038/s41392-023-01393-9

Figure Lengend Snippet: Fig. 4 AGR2-downregulation was an invasive biomarker in PCMR. a–d The AGR2 expression of BPH-versus tumor-derived primary cell in whole cell (a, b) and surface protein (c, d) lysates was evaluated employing LC-MS/MS (a, c) and western blotting (WB, b, d). The protein blot bands of WB were quantified utilizing gray scanning (ImageJ). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 by Wilcoxon rank-sum test. e IHC staining of AGR2 in different pathological grades (n = 138). The representative AGR2 IHC results (left) performed on Gleason score (GS) 3 + 4, GS 4 + 5, GS 5 + 5 and BPH. The positive rate of AGR2 was analyzed among BPHs (n = 37), early stage of tumors (GS 6–7, n = 80) and late stage of tumors (GS 8–10, n = 49) (right). *p < 0.05 by Fisher’s Exact test. Scale bar, 200 μm. f Kaplan–Meier plots for disease-free survival (DFS) of TCGA-PRAD patients grouped by the median of AGR2 levels. Statistics calculated by Log-rank test. g–i Scatter plots showing the Spearman’s correlation between AGR2 and three epithelium-related genes (CDH1, CTNNA1, CTNNB1) in TCGA PRAD (n = 494) database. The Spearman’s correlation was analyzed by GEPIA. j Spearman’s Correlation analysis between AGR2 and EMT-related genes (CDH1, CTNNA1, CTNNB1, EHF, VIM) in PCMR. k, l WB analysis of E-cadherin, Vimentin expression levels in NC/AGR2-KD PC3 cells (k) and xenograft tissues (l)

Article Snippet: After blocking membranes with 10% non-fat dried milk in phosphate-buffered saline and 0.1% Tween 20 solution, the following primary antibodies were incubated overnight at 4 °C with the membranes: Phospho-AKT (CST, 4060), AKT (CST, 4691), Phospho-c-MET (CST, 3077), c-MET (CST, 3127), Phospho-ALK (CST, 3983), ALK (CST, 3633), AMACR (Abcam, ab246927), AR (Abcam, ab268062), CK5 (Abcam, ab52635), E-cadherin (HuaAn, EM0502), Vimentin (HuaAn, EM0401), AGR2 (Proteintech, 12275-1- AP), Tubulin (Sigma-Aldrich, T6793), GAPDH (Proteintech, 10494-1- AP).

Techniques: Biomarker Discovery, Expressing, Derivative Assay, Liquid Chromatography with Mass Spectroscopy, Western Blot, Immunohistochemistry

Journal: Virology

Article Title: Impaired antiviral response in human hepatoma cells.

doi: 10.1006/viro.1999.9983

Figure Lengend Snippet: FIG. 7. Interferon-induced antiviral activity against VSV. Larynx epidermoid carcinoma Hep2, lung carcinoma A549 and hepatoma HepG2 and HuH7 cells were pretreated with different doses of IFN-a or IFN-g for 24 h followed by infection with VSV (1-5 pfu/cell). Virus-infected cells and cell culture supernatants were collected at 20 h after infection. (A) Western blot analysis. Blots (10 mg of protein/lane) were stained with rabbit anti-MxB (Hep2 cells only), anti-MxA, or anti-VSV antibodies as indicated in the figure. (B) Flow cytometric analysis. Cells were stained with rabbit anti-VSV antibodies and FITC-labeled goat anti-rabbit antibodies and analyzed by flow cytometry (FACScan with Cellquest software). Results are shown as relative viral protein expression. (C) Production of infectious VSV. Cell culture supernatants from IFN pretreated and virus-infected cells were subjected to virus plaque assay in Hep2 cells. Virus yields are shown in a logarithmic scale. IFN doses as indicated in the figure.

Article Snippet: HuH7 human hepatocellular arcinoma (Nakabayashi et al., 1982), Hep2 human larnx epidermoid carcinoma (ATCC CCL23), and A549 huan lung carcinoma (ATCC CCL 185) cells were mainained in minimal essential medium (MEM) with suppleents as described above. nterferons Type I human leukocyte IFN-a (6 3 106 IU/ml; specific ctivity of 2 3 108 IU/mg; Cantell et al., 1981) was kindly rovided by Dr. Kari Cantell at the National Public Health nstitute, Helsinki.

Techniques: Activity Assay, Infection, Virus, Cell Culture, Western Blot, Staining, Labeling, Flow Cytometry, Software, Expressing, Plaque Assay

Journal: Molecules

Article Title: 28-Hydroxy-3-oxoolean-12-en-29-oic Acid, a Triterpene Acid from Celastrus Orbiculatus Extract, Inhibits the Migration and Invasion of Human Gastric Cancer Cells In Vitro

doi: 10.3390/molecules24193513

Figure Lengend Snippet: ( A , C ) Changes in epithelial–mesenchymal transition (EMT) biomarker expression levels following 28-hydroxy-3-oxoolean-12-en-29-oic acid for 24 h were assessed by Western blotting. ( B , D ) The band intensities of E-cadherin, N-cadherin and vimentin relative to untreated control cells were quantified upon normalizing to β-actin expression, and are expressed as the mean ± standard deviation of three independent experiments.

Article Snippet: The reagents used included the following: RPMI 1640 cell culture medium, trypsin (HyClone, USA); fetal bovine serum (FBS) (Gibco, Waltham, MA, USA); artificially reconstituted basement membrane, Transwell chambers (Corning, Corning, NY, USA); thiazole blue (MTT) powder (Sigma, St. Louis, MO, USA); antibodies specific for E-cadherin, N-cadherin, vimentin, Snail, MMP-2, MMP-9, Akt, p-AKt, PI3K, p-PI3K, and β-actin (β-actin) (Cell Signaling, Danvers, MA, USA); and HRP-labeled goat anti-rabbit immunoglobulin (Ig) G (Hangzhou Huaan Biotechnology Co., Ltd., Hangzhou, China).

Techniques: Biomarker Discovery, Expressing, Western Blot, Control, Standard Deviation

Journal: Molecular cell

Article Title: Phosphorylated RB Promotes Cancer Immunity by Inhibiting NF-κB Activation and PD-L1 Expression

doi: 10.1016/j.molcel.2018.10.034

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Huamn: VCaP , ATCC , CRL-2876.

Techniques: Virus, Recombinant, Control, Mutagenesis, Kinase Assay, Sequencing, CRAfT Assay, Software