Metabolic Status Assessment in Cells

Complete RNA extraction from cell samples of all clusters was performed by using the TRIzol reagent (Thermo Fisher Scientific, Waltham, MA, USA) and then treated with a reverse-transcription kit (Thermo Fisher Scientific, Waltham, MA, USA). cDNA was quantified by quantitative PCR, using a QuantStudio 7 Flex Real-Time PCR system (Thermo Fisher Scientific, Waltham, MA, USA). The composition of the forward and reverse primers is shown in Table 2. A set of genes for assessing the metabolic status of cells was created on the basis of large studies of key metabolic pathways in different cancer types [21 (link)]. The results were calculated by the 2-ΔΔCT method, standardized for the U6 gene.