Journal: Cancers

Article Title: SNAT1 (SLC38A1) Is Not the Main Glutamine Transporter in Melanoma, but Controls Metabolism via Glutamine-Dependent Activation of P62 (SQSTM1)/cMYC-Axis

doi: 10.3390/cancers18071068

Figure Lengend Snippet: Validation of cMYC as a downstream effector of SNAT1 regarding ATP synthesis and mitochondrial respiration. ( A ) Exemplary measurement of OCR of Mel Im conducting the ATP rate assay. Melanoma cells were transfected with ZsGreen1-cMYC/pLVX-Puromycin or pLenti-C-GFP, respectively. ( B ) Calculation of ATP generated via glycolysis and oxidative phosphorylation, total ATP and basal respiration of Mel Im and Mel Juso. ATP production rate and OCR were normalized to 10,000 cells and compared to siCtr. Values represent the mean ± SEM of at least three independent experiments. The treatment and respective control group were compared using the Student’s unpaired t -test. p -value < 0.05 was considered statistically significant (*).

Article Snippet: All of the following functional experiments were conducted after a total transfection period of 96 h. Transfection with ZsGreen1-cMYC/pLVX-Puromycin vector was performed using 0.5 μg plasmid DNA and Lipofectamine LTX Plus reagent (Invitrogen, ThermoFisher Scientific) for a transfection period of 48 h. Andre Catic kindly gifted the ZsGreen1-cMYC/pLVX-Puromycin vector (RRID:Addgene_180278). pLenti-C-GFP (OriGene, Rockville, MD, USA; RRID:Addgene_17448) served as a control plasmid in our experiments.

Techniques: Biomarker Discovery, Transfection, Generated, Phospho-proteomics, Control