tissue lysates  (Novus Biologicals)

Journal: Scientific Reports

Article Title: Combination therapy of cisplatin with cilastatin enables an increased dose of cisplatin, enhancing its antitumor effect by suppression of nephrotoxicity

doi: 10.1038/s41598-020-80853-6

Figure Lengend Snippet: Megalin is not expressed in cancer cell lines. ( A ) Megalin expression in human lung cancer cell lines, a mouse colon adenocarcinoma cell line, human kidney lysate and mouse kidney lysate was determined by western blotting. The representative plots of 3 different experiments are shown. ( B ) mRNA expression of LRP2 (encoding megalin) in various cancers from the CCLE database. Normalized expression from the microarray data was calculated by robust multichip analysis (RMA).

Article Snippet: Whole human kidney lysate was obtained from Novus Biologicals (Littleton, CO, USA).

Techniques: Expressing, Western Blot, Microarray

Journal: Cell Death & Disease

Article Title: REGγ deficiency suppresses tumor progression via stabilizing CK1ε in renal cell carcinoma

doi: 10.1038/s41419-018-0646-2

Figure Lengend Snippet: a , b Expression of REGγ in RCC tissues (T) and normal kidney tissues (N) as detected by IHC ( a ) and WB ( b ). c Expression of REGγ in four RCC cell lines (A498, 786-O, ACHN, and caki-1) and the normal renal tubular epithelial cell line (HK-2) as detected by western blot. d Comparison of REGγ expression in different Fuhrman grades I–IV of RCC tissue samples via IHC staining (left panel: magnification ×200, scale bar = 50 μm; right panel: magnification ×400, scale bar = 20 μm). e REGγ (PSME3) expression (median of expression intensity) in different pathological types and grades of RCC derived from Oncomine database ( https://www.oncomine.org/ ). f Kaplan–Meier analysis of the correlation between REGγ expression and the survival time in RCC patients. Cases were classified into lower expression group and higher expression group as described in methods. g Prognosis of RCC (KIRC) patients with high or low expression of REGγ (PSME3) derived from OncoLnc database ( https://www.oncolnc.org/ )

Article Snippet: The stable REGγ knockdown RCC cell lines ACHN and A498 were generated by integration of retroviral shREGγ vectors specific for REGγ or a control gene (GFP) from OriGene (Rockville, MD), which was performed as previously described .

Techniques: Expressing, Western Blot, Immunohistochemistry, Derivative Assay

Journal: Cell Death & Disease

Article Title: REGγ deficiency suppresses tumor progression via stabilizing CK1ε in renal cell carcinoma

doi: 10.1038/s41419-018-0646-2

Figure Lengend Snippet: a PSME3 upregulation is closely correlated with renal cell carcinoma revealed by KEGG pathway analysis. b GSEA indicated that cell proliferation and anti-apoptosis were positively correlated with elevated PSME3 expression in RCC from database GSE89563 (GO_0008284 and GO_0006915). NES, normalized enrichment score. c Stable knockdown of REGγ (shREGγ) in RCC cell lines (ACHN and A498) confirmed by WB. d , e Effect of shREGγ on RCC cells growth as determined by MTT assay. f Representative images of colony formation of RCC cells after transfection of shREGγ versus shNC. g , h Representative images of EdU incorporation assay after transfection of shREGγ versus shNC. i , j Representative flow cytometry plots of cell cycle distribution from ACHN and A498 cells transfected with shREGγ and shNC. k , l Apoptosis rate from ACHN and A498 cells after transfected with shREGγ and shNC as detected by flow cytometry. Data are shown as mean ± SD. * P < 0.05

Article Snippet: The stable REGγ knockdown RCC cell lines ACHN and A498 were generated by integration of retroviral shREGγ vectors specific for REGγ or a control gene (GFP) from OriGene (Rockville, MD), which was performed as previously described .

Techniques: Expressing, MTT Assay, Transfection, Flow Cytometry

Journal: Cell Death & Disease

Article Title: REGγ deficiency suppresses tumor progression via stabilizing CK1ε in renal cell carcinoma

doi: 10.1038/s41419-018-0646-2

Figure Lengend Snippet: a , b Representative images and the relative quantification of wound-healing assay in RCC cells transfected with shREGγ and shNC. c , d Representative images and the relative quantification of transwell invasion assay in ACHN and A498 cells transfected with shREGγ and shNC. Data are shown as mean ± SD. * P < 0.05

Article Snippet: The stable REGγ knockdown RCC cell lines ACHN and A498 were generated by integration of retroviral shREGγ vectors specific for REGγ or a control gene (GFP) from OriGene (Rockville, MD), which was performed as previously described .

Techniques: Wound Healing Assay, Transfection, Transwell Invasion Assay