Fig. 5: Hormone-mediated ATM activation impacts SIM2s function.
From: SIM2s directed Parkin-mediated mitophagy promotes mammary epithelial cell differentiation
A Expression of activated (phosphorylated) ATM in HC11 cells at an undifferentiated state and 24 h after adding hydrocortisone alone, prolactin alone, or hydrocortisone + prolactin. B ATM consensus phosphorylation sites on SIM2s. C Co-immunoprecipitation of ATM and FLAG in Sim2s or Sim2s-pATMΔ HC11 cells at 24 h differentiation. D mRNA expression of differentiation-dependent Csn2 in control, Sim2s, and Sim2s-pATMΔ HC11 cell lines (n = 2 independent experiments with biological triplicates and technical duplicates). E Protein levels of MAP1LC3B I and II in control and Sim2s-pATMΔ HC11 across differentiation. F Basal energy phenotype in control, Sim2s, and Sim2s-pATMΔ HC11 cells across differentiation (n = 2 independent experiments with ≥10 technical replicates each). G Proposed model of ATM-mediated phosphorylation of SIM2s and its downstream effects. bHLH basic helix-loop-helix, ECAR extracellular acidification rate, H hydrocortisone, NLS nuclear localization sequence, OCR oxygen consumption rate, P prolactin, PAS PER-ARNT-SIM, U undifferentiated. Error bars indicate the mean ± SD. **p < 0.01.
