Genetic Manipulation of ZNF750 in Cells

Expression vectors pLEX-ZNF750 (wildtype) and pLEX-C2H2 mutant ZNF750 (C2H2 mutant) were generously provided by Dr Paul A. Khavari (Stanford University School of Medicine). We also used another ZNF750 expression vector, SHC003-ZNF750 (which we reported previously^{3 (link)}), in some of the experiments to confirm that the results were not affected by different vector systems (Supplementary Figure 6). The shRNA construct for p63 was made with PLKO.1 backbone (Sigma-Aldrich) using Age I and EcoR I sites (Supplementary Table S3). LAMC2 over-expression and shRNA vectors were described previously.²⁴ Pooled siRNAs targeting human ZNF750 (M-014417-01) and TINCR (R-015725-00-0005) and scramble siRNA (D-001210-01) were purchased from Thermo Scientific (Rockford, IL, USA) (Supplementary Table S3).

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Hazawa M., Lin D.C., Handral H., Xu L., Chen Y., Jiang Y.Y., Mayakonda A., Ding L.W., Meng X., Sharma A., Samuel S., Movahednia M.M., Wong R.W., Yang H., Tong C, & Koeffler H.P. (2016). ZNF750 is a lineage-specific tumour suppressor in squamous cell carcinoma. Oncogene, 36(16), 2243-2254.

Publication 2016

PLKO.1 backbone

Backbone Human Shrna Sirna Vectors

Corresponding Organization : National University of Singapore

Other organizations : Kanazawa University, University of California, Los Angeles, Cedars-Sinai Medical Center

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Variable analysis

independent variables

Expression vectors pLEX-ZNF750 (wildtype)
Expression vector pLEX-C2H2 mutant ZNF750 (C2H2 mutant)
Expression vector SHC003-ZNF750
ShRNA construct for p63 with PLKO.1 backbone
LAMC2 over-expression vectors
LAMC2 shRNA vectors
Pooled siRNAs targeting human ZNF750
Pooled siRNAs targeting TINCR
Scramble siRNA

dependent variables

Outcomes measured in the experiments

control variables

Scramble siRNA

positive controls

SHC003-ZNF750 expression vector

negative controls

Scramble siRNA

Annotations

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