Figure S4 from Arginine Depletion Therapy with ADI-PEG20 Limits Tumor Growth in Argininosuccinate Synthase–Deficient Ovarian Cancer, Including Small-Cell Carcinoma of the Ovary, Hypercalcemic Type

Abstract

ASS1 in ovarian cancer cell lines. A, Re-expression of SMARCA4 does not affect the expression of ASS1. SCCOHT cells were infected with Lenti-GFP or Lenti-SMARCA4 for 24 hours followed by puromycin selection for 72 hours. Cells were then harvested for protein quantification using mass spectrometry. B, ASS1 immunohistochemistry on FFPE cell line pellets showing positivity in JHOC 7 and JHOC 9, and lack of expression in ES2, JHOC 5, and TOV112D. C, ASS1 immunohistochemistry on FFPE cell line pellets for CRISPR knock out (JHOC 7 ASS1 CRISPR A8) and over-expression (JHOC 5 ASS1 CMV) cell lines compared to their corresponding vector controls (JHOC 7 CRISPR NTC30 and JHOC5 vector control, respectively). D, qPCR showing differential ASS1 mRNA expression levels between ovarian cancer cell lines. Difference of relative mRNA expression between cell lines is determined by one-way ANOVA with post-hoc Tukey test. * p<0.05 when compared to JHOC 7; ## p <0.001 when compared to JHOC 9. Error bars represent standard error of mean. E, ASS1 promoter methylation status, all samples were run on the same gel. F, Cell lines are sensitive to depletion of arginine and citrulline from growth media. Addition of citrulline rescued the growth of ASS1 expressing JHOC7, but not the ASS1 deficient JHOC 5 and COV434. -