Supplemental Methods, Figures 1 - 12, Tables 1 - 2, 4 from A Chemical Genetics Approach for the Functional Assessment of Novel Cancer Genes

Autor: Frank Stegmeier, Michael R. Schlabach, Junxia Min, Yi Yang, HoMan Chan, Veronica Gibaja, Michelle Lira, Iris Kao, Daniel Rakiec, David Ruddy, Adnan Derti, Qianhe Zhou
Rok vydání: 2023
DOI: 10.1158/0008-5472.22405304.v1
Popis: Supplemental Figure 1. Efficient knock�in of DD�domain into the endogenous p53 locus by TALENs in HCT116 cells. Supplemental Figure 2. PCR verification of Degron�KI clones. Supplemental Figure 3. Immunofluorescence staining of Degron�KI cells. Supplemental Figure 4. Quantification of DD tagged proteins in the presence or absence of Shld. Supplemental Figure 5. Highly efficient Degron�KI at the endogenous EZH2 locus by CRISPR in HCT116. Supplemental Figure 6. Assessment of the kinetics of DD�EZH2 and DD�SF3B1 depletion upon Shld withdrawal. Supplemental Figure 7. HCT116 (EZH2 wild type) cells are not sensitive to EZH2 inhibitor EI1. Supplemental Figure 8. RT�PCR strategy to identify allele� specific DD tagging of mutant versus wildtype SF3B1. Supplemental Figure 9. UQCC and CRNDE exhibit an altered splice pattern in SF3B1 mutant uveal melanoma cell lines. Supplemental Figure 10. Shld has minimal effects on gene expression of parental Mel202 cells and Mel202 DD�mut�SF3B1. Supplemental Figure 11. Selective depletion of mutant SF3B1 in Mel202 Degron�KI cells reversed the alternative splicing pattern of DYNLL1, SNRPN, TMEM14C, ABCC5, ZDHHC16, RBM18. Supplemental Figure 12. Selective depletion of mutant SF3B1 in Mel202 Degron�KI cells by Shld withdrawal predominantly alters 3' splice sites but not 5' splice site selection. Supplemental Figure 13. Confirmation that the DD tag insertion occurred exclusively at the SF3B1 locus in Degron�KI engineered Mel202 clones. Supplemental Table 1: Detailed genotypes of ESS�1 Degron�KI clones. Supplemental Table 2: Detailed genotypes of Mel202 Degron�KI clones. depletion using MATS. Supplemental Table 4: Comparison of the alternatively spliced genes identified in this study with genes reported in prior reports (5).
Databáze: OpenAIRE