Fig. 6
Gene expression changes in MAPK/ERK-deficient nephron progenitors suggest defects in mitochondrial functions. A Further heatmap analysis on the MM dataset revealed that mitochondria-related biological processes are affected in the absence of MAPK/ERK activation. The identified genes are listed in Additional file 9: Table S8. B Mitochondrial DNA copy number analysis was performed on control (DMSO, n = 9) and MEK1/2-inhibited (U0126, n = 9) E12.5 kidneys by real-time PCR. Mitochondrial DNA was measured by the analysis of its 12S expression against nuclear DNA quantification by Rbm expression. C Quantification of ATP from mK4 cell line derived from embryonic kidney mesenchyme by LC/MS (n = 4 replicates for each DMSO and U0126). ATP concentrations are normalized against total protein concentration (Additional file 10: Table S9). **p < 5 × 10−3, ***p < 5 × 10−6. D Oxygen consumption rate (OCR) of mK4 cell line was measured by a Seahorse XF analyzer. For the OCR measuring, ATP synthase inhibitor (oligomycin), protonophore uncoupler (FCCP), and ETC inhibitors (rotenone and antimycin A) were added at the indicated points (n = 4 replicates for each DMSO and U0126). E Basal respiration, ATP production, proton leak, maximal respiration, and spare capacity measures are shown in different samples. Error bars represent standard deviation (S.D.). NT, non-treated mK4 cells; DMSO, DMSO-treated mK4 cells as a control; U0126, MEK inhibitor U0126-treated mK4 cells