(A) Schematic illustrating the global phosphoproteomics experiment. (B) The heatmap depicts phosphopeptides that are either positively or negatively correlated with the transformation phenotype (p<0.… see more
(A) PP2A Ca after knockdown using shPP2A Ca1 and shPP2A Ca2 and (B) PP2A B56g after knockdown using shPP2A B56g1 and shPP2A B56g2 as measured by RNAseq (Reads Per Kilobase of transcript, per Million … see more
(A) Schematic of experimental design to reveal binding proteins that when depleted can substitute for ST in transformation (B) Representative image of AI growth induced by ST or MAP4K4 partial … see more
Quantification of soft-agar colony and tumor volume with MAP4K4 knockdown.
(A) AI colony count of HEK TER cells expressing the corresponding shRNAs or SV40 ST. Student’s t-test was performed for each shRNA compared to shLuc control. (B) AI colony count of HEK TER cells … see more
(A) Schematic of targeted proteomic analysis of MAP4K4 phosphorylation and interacting proteins in the presence of ST or GFP control. (B) SILAC experiment in which MAP4K4-associated proteins were … see more
Quantification of MAP4K4 interacting proteins and phosphopeptides.
(A) Co-immunoprecipitation analysis in 293T cells of CTAP-tagged SV40 ST wild-type and PP2A binding mutants. Co-immunoprecipitation of STRN3 and Aα is shown. (B) Co-immunoprecipitation analysis of … see more
(A) Quantification of MAP4K4 in vitro kinase activity after MAP4K4 was tandem-affinity purified from cells expressing ST or GFP control. (B) Quantification of AI growth after increasing … see more
Quantification of AI growth and in vitro MAP4K4 kinase activity.
(A) In vitro kinase assay of tandem-affinity purified MAP4K4 (Flag/HA) from HEK TER cells expressing either ST or GFP was performed and MBP was used as a substrate. HA and phospho-MBP were detected … see more
(A) Schematic of experimental design to reveal binding proteins, that when depleted, inhibit ST-mediated transformation. (B) Representative images of AI colonies observed after knockdown of STRN4 in … see more
Quantification of soft-agar colony and tumor volume with STRN4 knockdown.
(A) AI colony count of HEK TER ST expressing four different STRN4 shRNAs or shLuc control. Immunoblot below depicts corresponding STRN4 expression after the knockdown. (B) Correlation between the … see more
(A) Schematic of proteomic analysis of STRN4 interacting proteins in the presence of ST or a GFP control. (B) Fold change in abundance of STRN4 interacting proteins in the presence of ST compared to … see more
Qunatification of STRN4 interacting proteins and in vitro MAP4K4 kinase activity and AI growth.
(A) Detection of ST from co-immunoprecipitation of STRN4 GFP- and ST-expressing cells through proteomic analysis. The values were normalized to STRN4 counts. ST was not detected (N.D.) in the … see more
(A) Heatmap of Enrichment Scores (ES) from RNA-seq analysis showing that partial suppression of MAP4K4 expression in HEK TER cells upregulates a transcriptional signature closely resembling four … see more
Quantification of CTGF and CYR61 gene expression (TPM).
(A) Quantification of AI growth obtained following partial MAP4K4 suppression alone or when combined with YAP1 suppression (shYAP1) in HEK TER cells. Transformation induced by partial MAP4K4 … see more
Quantification of AI growth with changes in YAP1 and MAP4K4.
(A) Immunoblot depicting changes in MAP4K4 and YAP1 protein levels upon either depleting YAP1 alone, or in combination with MAP4K4. (B) Immunoblot depicting the overexpression of YAP1 WT or YAP1 S5A … see more
Key Resources Table.
Normalized iTRAQ phosphoproteomic profiles of changes in phosphopetides upon suppression of PP2A Cα, Aα, B56γ or SV40ST expression.
Results of the SILAC experiment representing MAP4K4 interacting proteins.
Results of the SILAC experiment representing targeted MAP4K4 phospho-profiling.
Results of MudPIT experiment showing STRN4 interacting proteins.
RNAseq (TPM) profiles of MAP4K4 knockdown (shMAP4K4-82).
Genesets used in the study.