Fig. 6
ADSL controls cMYC negative regulator MIR22HG expression. a Scatterplot of differential-expression results: −log10 FDR-adjusted P-values from comparisons of each gRNA versus control using DESeq2 are plotted for all genes regardless of significance. b, c MIR22HG mRNA expression upon ADSL depletion in (b) MDA-MB-231 and (c) MDA-MB-436 cells. Graphs represent the mean ± SEM, n = 3 (b) and n = 4 (c). d Pearson correlation between the expression of ADSL and MIR22HG in TNBC patients from TCGA dataset. e MIR22HG mRNA expression and f cMYC protein level after transfecting MDA-MB-231 cells with either MIR22HG expressing plasmid or empty vector. g MIR22HG mRNA expression and h cMYC protein level after transfecting MDA-MB-231 cells with either three independent siRNAs targeting MIR22HG or siRNA control. i MIR22HG mRNA expression, j 2-D, and k 3-D colony formation in the presence or absence of doxycycline (dox) in MDA-MB-231 cells transduced with dox-inducible MIR22HG expressing lentivirus. Graphs represent the mean ± SEM from four independent sets of samples (i), and from two independent experiments, each performed in duplicate (k). l, m MIR22HG mRNA expression in ADSL control or knockout MDA-MB-231 cells with or without the expression of (l) WT and (m) P24A ADSL. n MIR22HG mRNA expression in ADSL control or knockout MDA-MB-231 cells treated as indicated (adenosine concentration was 50 µM). Graphs in (l), (m), and (n) represent the mean ± SEM, n = 3. o Schematic of the proposed mechanism by which EglN2-hydroxylated ADSL controls cMYC and cMYC target gene expression. *P < 0.05, **P < 0.01, ***P < 0.001 were calculated using one-way ANOVA followed by Dunnett’s multiple comparison test in (b), (c), and (g). In (e), (i), and (k), P-values were calculated using two-tailed Student’s t-test. In (l), (m), and (n), P-values were calculated using one-way ANOVA followed by Tukey’s multiple comparison test. Source data are provided as a Source Data file
