Journal: Biomedicines
Article Title: RICTOR Affects Melanoma Tumorigenesis and Its Resistance to Targeted Therapy
doi: 10.3390/biomedicines9101498
Figure Lengend Snippet: RICTOR-mediated melanoma spheroid growth induces an MIC phenotype ( A , B ) Colo829, A375, and SkMel5 cell lines were cultivated under the same condition as and treated with DMSO or an mTOR inhibitor (OSI-027 1 μM) for 10 days and counted (scale bar represents 1000 μm). Graphs represent the mean ± s.d. of an experiment in triplicate (similar results were obtained in three independent experiments). Significance was calculated using the Student’s t -test: * p < 0.05, ** p < 0.01. ( C ) The expressions of Myc-tagged RICTOR, RICTOR, phosphorylated STAT3 (Y705), total STAT3, MITF, SOX10, and AXL were assessed by Western blotting in Colo829 cells transfected with an empty vector (Control) or with a vector expressing human RICTOR cDNA (RICTOR). ( D ) Relative mRNA expressions of the different tumor-initiating cell markers (JARID, Tie1, and BMPR1), as well as RICTOR, were assessed by qPCR and normalized to GAPDH in Colo829 cells transfected with an empty vector (Control) or with a vector expressing human RICTOR cDNA (RICTOR).
Article Snippet: Membranes were probed with the following primary antibodies: Myc-Tag, RICTOR, ACTIN, p-AKT (S473), AKT, p-ERK, ERK, pSTAT3 (Y705), STAT3, SOX10, AXL (Cell Signaling Technology, Danvers, MA, USA), and MITF (OriGene Technologies Inc, Rockville, MD, USA).
Techniques: Western Blot, Transfection, Plasmid Preparation, Control, Expressing