Fig. 7
FOXH1 is an additional activator of autophagy and responds to the HDACs/MYC axis. a Motif analysis using HDAC2-binding sites for autophagy genes identified the FOXH1 motif. b RT-qPCR of autophagy genes in FOXH1-overexpressing HeLa cells (n = 6 biologically independent samples) p < 0.01. c Representative LC3 immunoblot from FOXH1-overexpressing cells. d Quantification of the LC3II/LC3I ratio normalized to Coomassie stained immunoblots, used as loading control (n = 6 independent experiments). e FOXH1 RT-qPCR in SAHA-treated HeLa cells (20 μM for 24 h; n = 7 biologically independent samples). f Representative immunoblot of FOXH1 overexpression in HeLa cells treated with DMSO or SAHA (20 μM for 24 h). g Quantification of f (n = 3 independent experiments) normalized to Coomassie stained immunoblots, used as loading control. h MYC binding to the FOXH1 locus obtained from ChIP-seq datasets for U2OS cells overexpressing MYC. i Expression of the FOXH1 gene in U2OS cells with MYC overexpression, data obtained from Walz et al. j RT-qPCR of Foxh1 in tumorpheresTrp53−/− overexpressing Myc. Values are relative to those obtained in control Trp53−/− cells (n = 5 biologically independent samples). k RT-qPCR of FOXH1 in U2OS cells in which MYC was silenced by shRNA. Boxes represent the mean value and bar inside the box represents median value; upper bar represents maximum of distribution; lower bar represents minimum of distribution (95% confidence level). Graphs are presented as mean ± SD. Statistical analysis was performed using the Student t-test
