Skip to main content
. 2021 Mar 26;12:1920. doi: 10.1038/s41467-021-22101-7

Fig. 3. AAMDC regulates targets involved in cell proliferation and metabolism.

Fig. 3

a Gene ontology (GO) analyses of the top 10 biological processes (sorted by Modified Fisher Exact p-value, EASE score by DAVID) identified by analysis of differentially regulated genes as assessed by RNA-seq of SUM52PE cells transduced with AAMDC shRNA #2 (sh2) compared with empty vector (EV) (Supplementary Data 2). b Network analysis (STRING, v11) outlining interactions between differentially regulated targets. MTHFD1L is outlined in red. c Schematic representation of 1C metabolism (folate and methionine cycles) and amino acid biosynthesis (serine and asparagine) pathways. Genes regulated by AAMDC (as per RNA-seq) are depicted in dark blue. 3PHP 3-phosphohydroxypyruvate, 3PS 3-phosphoserine, 3PG 3-phosphoglycerate, THF tetrahydrofolate, TCA tricarboxylic acid cycle. d Representative immunoblots (IB) showing the reduction of MTHFD1L protein expression by the AAMDC knockdowns (KDs) shown in Fig. 2a. Source data are provided as a Source Data file. e Knockdown of MTHFD1L by shRNA decreased cell proliferation in SUM52PE cells. Protein levels are assessed by IB (top) and cell proliferation by α-Ki-67 staining (green); Hoechst 33258-stained nuclei (blue). WT wild-type (untransduced cells), EV empty vector, MTHFD1L shRNAs #1–#2 (sh1–2). Representative images are shown (middle). Quantification is shown in bottom panel: WT (gray), EV (blue), shRNA #1 (orange), shRNA #2 (red). The relative quantification of Ki-67+ cells is normalized to EV and presented as mean values ± SD. p-values are determined by two-tailed unpaired t-test (**p = 0.0039, ****p < 0.0001). Fields of view examined: n = 7 for EV, n = 9 for WT and sh1, and n = 10 for sh2 over 2 independent experiments (right panel). f The top 25 significantly differentially regulated metabolites (WT vs. AAMDC sh2; p < 0.05, two-tailed unpaired t-test) identified by liquid chromatography-mass spectrometry (LC-MS) in SUM52PE cells stably transduced with either AAMDC sh2 or EV. n = 3 biologically independent experiments.