Fig. 2

Defining the MTHFD2 interactome. a Schematic of the strategy used for identifying MTHFD2 interactors. Immunoprecipitation was performed on parental HCT-116 cells (WT) and MTHFD2 knockout cells (D2-KO) using either one of two anti-MTHFD2 antibodies (AP and NC) or an anti-rabbit immunoglobulin (IgG) antibody. The numbers above antibody names indicate the number of replicate experiments performed for a given condition. After in-gel digestion and MS 29 proteins were found to be detected in IP from both anti-MTHFD2 antibodies in parental HCT-116 cells but not in IP performed in MTHFD2 knockout cells or using IgG antibody. b Representative silver-stained SDS-gel containing co-immunoprecipitated samples. Lane 1 contained a molecular weight ladder. Lane 2 and 3 lysates from wildtype HCT-116 cells after immunoprecipitation with anti-MTHFD2 antibody, lanes 4 and 5 lysates from same cells immunoprecipitated using anti-rabbit IgG antibody. c Number of unique peptides matching to MTHFD2 identified in replicate experiments with two anti-MTHFD2 antibodies in WT or knockout D2-KO cells. d, e Selected significantly enriched GO Molecular Function and Cellular Component respectively. f PPI network generated for 29 identified MTHFD2 interactors using experimental evidence in STRING database and medium confidence links (> 0.400). Nodes with no interaction are not shown. Thickness of edges represent strength of evidence supporting interaction. Members of protein functional groups are indicated according to color chart