Fig. S4 from Nuclear PTEN Localization Contributes to DNA Damage Response in Endometrial Adenocarcinoma and Could Have a Diagnostic Benefit for Therapeutic Management of the Disease

Supplementary Figure 4. Effects of Zeocin treatment on levels of �H2AX and accumulation of cells in G2/M phase in PTEN-null Ishikawa cells after the overexpression of four different PTEN plasmids. Cells were transfected with plasmids for 24 h, treated with Zeocin for 16 h, then assayed. A-P Levles of �H2AX as determined by co-immunocytostaining with anti-PTEN antibody followed by Alexa Fluor 488 conjugated secondary antibody (green) and anti-phospho H2AX (Ser 139) antibody followed by Alexa Fluor 546 conjugated secondary antibody (red). Nuclei were stained with DAPI (blue). A-D, pEGFP-EV. E-H, PTEN-WT. I-L, PTEN-NLS. M-P, PTEN-NES. Scale bar; 200 μm. Q, Comparison of % �H2AX+ cells as determined by flow cytometry analysis in PTEN transfected cells with or without Zeocin treatment. R, Comparison of the percentage of cells in G2/M as determined by flow cytometry analysis in PTEN transfected cells with or without Zeocin treatment. Statistical analysis performed was paired T-test for each plasmid P<0.05</p

Fig. S1 from Nuclear PTEN Localization Contributes to DNA Damage Response in Endometrial Adenocarcinoma and Could Have a Diagnostic Benefit for Therapeutic Management of the Disease

Supplementary Figure 1. A, Oncoprints of PTEN and TP53 in four molecular subtypes of EndoCA based on TCGA. B, Mutational landscape of PTEN in EndoCA. DSPc; dual specificity phosphatase domain, C2; cell membrane binding domain. Results were generated using C-BioPortal website algorithms (48, 49).</p

Fig. S3 from Nuclear PTEN Localization Contributes to DNA Damage Response in Endometrial Adenocarcinoma and Could Have a Diagnostic Benefit for Therapeutic Management of the Disease

Supplementary Figure 3. PTEN localization in cytoplasm and nuclei of epithelial cells in ANT and EndoCA by grade. PTEN immunostaining was performed on adjacent normal tissues (ANT) and matched primary endometrial tumors (EndoCA). Quantification of staining intensities was assessed by histo-scoring (H-score) in cytoplasm and nuclei of epithelial cells. Average intensities are expressed as median with 95% CI, and statistical significance are determined by Wilcoxon matched-pairs signed rank test. A, n=10 matched ANT and Grade1 tumors. B, n=9 matched ANT and Grade 2 tumors. C, n=10 matched ANT and Grade 3 tumors.</p

Fig. S2 from Nuclear PTEN Localization Contributes to DNA Damage Response in Endometrial Adenocarcinoma and Could Have a Diagnostic Benefit for Therapeutic Management of the Disease

Supplementary Figure 2. A and B, PTEN immunostaining in adjacent normal tissue (ANT) and EndoCA, respectively. C and D, Immunostaining of phospho-AKT (Ser473) in ANT and EndoCA, respectively. E, TCGA analysis of PTEN expression in human endometrial tumors by high throughput reverse phase protein lysate microarray (RPPA). F-H, PTEN immunostaining of human normal endometrial tissue with the anti-human PTEN antibody generated from three different clones. EG; endometrial gland. Str; stroma.</p