Journal: Open Medicine

Article Title: Lathyrol affects the expression of AR and PSA and inhibits the malignant behavior of RCC cells

doi: 10.1515/med-2024-1136

Figure Lengend Snippet: Primary antibody dilutions for WB

Article Snippet: The following reagents and materials were used in this study: 786-O human RCC cell line authenticated by RTS (Short Tandem Repeat) authentication and mycoplasma tests (Procell Life Science & Technology Co., CL-0010); lathyrol (Weikeqi-Biotech, Sichuan, China, Wakq-00424); nilutamide (AR blocker; Abmole Biotechnology Co., Ltd); fetal bovine serum (FBS; Absin Bioscience); CCK-8 reagent and phosphate buffer saline (PBS) buffer (Biosharp); 1640 basal medium, trypsin-ethylene diamine tetraacetic acid (0.25%) digestion solution, cell culture-grade dimethylsulfoxide (DMSO), tissue and cell radio-immunoprecipitation assay (RIPA) lysis buffer, phenylmethanesulfonyl fluoride, goat serum, hydrogen peroxide (H 2 O 2 ), Mayer’s hematoxylin staining solution, crystal violet staining solution, and neutral balsam (Solarbio); enhanced chemiluminescence (ECL) developing solution, horse radish peroxidase (HRP)-labeled sheep anti-rabbit secondary antibody for immunohistochemistry, and diaminobenzidine (DAB) chromogenic kit (Servicebio); bicinchoninic acid assay (BCA) protein quantification kit, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) cell apoptosis detection kit (green fluorescence), Triton X-100 (cell permeabilization agent), 4′,6-diamidino-2-phenylindole nuclear stain, mounting medium with anti-fluorescence quenching reagent, and HRP-labeled goat anti-rabbit secondary antibody (Beyotime Biotechnology); 6-well Transwell plates and Matrigel (Corning Biotechnology); rabbit polyclonal anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody (internal control; Hangzhou Xianzhi Biotechnology); rabbit polyclonal anti-Ki67 antibody (Bioss Biotechnology); 4% paraformaldehyde fixative solution and anti-phosphorylated adrenergic receptor (p-AR) antibody (phosphorylation site, S650; Absin Biotechnology); protein marker (10–180 kD), anti-phosphorylated Akt (p-Akt) antibody (phosphorylation site, Ser473), rabbit polyclonal anti-Bax antibody, anti-Bcl-2 antibody, anti-matrix metalloproteinase 2 (MMP2) antibody, anti-MMP9 antibody, and anti-cleaved-caspase3 & caspase3 antibody (Wuhan Sanying Biotechnology); rabbit polyclonal anti-AR antibody and rabbit monoclonal anti-cleaved-caspase9 & caspase9 antibody (Affinity Biotechnology); and rabbit monoclonal anti-PSA antibody (STARTER Biotechnology).

Techniques:

Journal: Open Medicine

Article Title: Lathyrol affects the expression of AR and PSA and inhibits the malignant behavior of RCC cells

doi: 10.1515/med-2024-1136

Figure Lengend Snippet: (a) Western blot results for Ki67 and GAPDH protein expression, (b) statistical analysis of Ki67 expression levels, and (c) statistical analysis of the CCK-8 results. The numerical details can be seen in .

Article Snippet: The following reagents and materials were used in this study: 786-O human RCC cell line authenticated by RTS (Short Tandem Repeat) authentication and mycoplasma tests (Procell Life Science & Technology Co., CL-0010); lathyrol (Weikeqi-Biotech, Sichuan, China, Wakq-00424); nilutamide (AR blocker; Abmole Biotechnology Co., Ltd); fetal bovine serum (FBS; Absin Bioscience); CCK-8 reagent and phosphate buffer saline (PBS) buffer (Biosharp); 1640 basal medium, trypsin-ethylene diamine tetraacetic acid (0.25%) digestion solution, cell culture-grade dimethylsulfoxide (DMSO), tissue and cell radio-immunoprecipitation assay (RIPA) lysis buffer, phenylmethanesulfonyl fluoride, goat serum, hydrogen peroxide (H 2 O 2 ), Mayer’s hematoxylin staining solution, crystal violet staining solution, and neutral balsam (Solarbio); enhanced chemiluminescence (ECL) developing solution, horse radish peroxidase (HRP)-labeled sheep anti-rabbit secondary antibody for immunohistochemistry, and diaminobenzidine (DAB) chromogenic kit (Servicebio); bicinchoninic acid assay (BCA) protein quantification kit, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) cell apoptosis detection kit (green fluorescence), Triton X-100 (cell permeabilization agent), 4′,6-diamidino-2-phenylindole nuclear stain, mounting medium with anti-fluorescence quenching reagent, and HRP-labeled goat anti-rabbit secondary antibody (Beyotime Biotechnology); 6-well Transwell plates and Matrigel (Corning Biotechnology); rabbit polyclonal anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody (internal control; Hangzhou Xianzhi Biotechnology); rabbit polyclonal anti-Ki67 antibody (Bioss Biotechnology); 4% paraformaldehyde fixative solution and anti-phosphorylated adrenergic receptor (p-AR) antibody (phosphorylation site, S650; Absin Biotechnology); protein marker (10–180 kD), anti-phosphorylated Akt (p-Akt) antibody (phosphorylation site, Ser473), rabbit polyclonal anti-Bax antibody, anti-Bcl-2 antibody, anti-matrix metalloproteinase 2 (MMP2) antibody, anti-MMP9 antibody, and anti-cleaved-caspase3 & caspase3 antibody (Wuhan Sanying Biotechnology); rabbit polyclonal anti-AR antibody and rabbit monoclonal anti-cleaved-caspase9 & caspase9 antibody (Affinity Biotechnology); and rabbit monoclonal anti-PSA antibody (STARTER Biotechnology).

Techniques: Western Blot, Expressing, CCK-8 Assay