IL-6 knockout suppresses proliferation and migration and induces a metabolic quiescent state in TSC2-deficient cells. (A) Secreted IL-6 is decreased in three IL-6 CRISPR/Cas9 clones compared to control, as measured by ELISA. (B) IL-6 knockout decreases the proliferation of TSC2-deficient MEFs compared to control as measured by crystal violet staining as a readout of cell density. (C) Representative images of (D) quantified cells migrated through transwells toward serum, which was decreased in IL-6 knockout; TSC2-deficient MEFs compared to TSC2-deficient control MEFs. (Scale bar, 100 μm.) (E) OCR is decreased in TSC2-deficient MEFs with IL-6 knockout compared to control cells. Data show measurements from the Seahorse extracellular flux analyzer using the MitoStress assay. (F) Summary and statistical analysis of OCR results. (G) Basal ECAR is decreased in TSC2-deficient MEFs with IL-6 knockout MEFs compared to control. mpH, milli-pH. (H) Energy map showing the global bioenergetic status of TSC2-deficient MEFs with IL-6 knockout compared to control. Data presented as the mean ± SD of three to four independent experiments. One-way ANOVA and Student’s t test were used for statistical analysis. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.