Immunoblotting Procedures and Antibodies

Immunoblotting was conducted as previously described (15 (link),25 (link)). The primary antibodies used for western blot were c-Myc (SC-40, 1:500, Santa Cruz Technology); phospho-c-Myc (#13748, c-Myc-pSer62, 1:500; Cell Signaling Technology); phospho-c-Myc (Y011034, c-Myc-pThr58, 1:1000; Applied Biological Materials Inc.); RPA2 (Clone NA18, 1:100, Calbiochem/EMD Millipore); anti-β-actin (Clone AC-74, 1:50000, Sigma-Aldrich); anti-CHK1 (G-4, 1:200, Santa Cruz Technology); phospho-CHK1 antibody (#133D3, CHK1-pSer345, 1:500, Cell Signaling Technology); CDC45 (G-12 sc55569, 1:200, Santa Cruz Technology); γ-H2Ax (ser139, clone JBC301, 1:500, Millipore); rabbit polyclonal antibody phosphor RPA32 Ser4/Ser8 (BL647, 1: 1000, Bethyl; Histone H3 (#9715, 1:1000, Cell Signaling Technology); PPP2R5A (ab89621,1:1000, Abcam); eIF4E (A2162, 1:1000, Abclonal); Phospho-eIF4E (#9741, eIF4E-Ser209, 1:1000, Cell Signaling Technology); 4E-BP1 (#9452, 1:1000, Cell Signaling Technology); Phospho-4E-BP1 (#2855, 4E-BP1-Thr36/47, 1:1000, Cell Signaling Technology); Phospho-p70S6K antibody (#9205, p70S6K-Thr389, 1:1000, Cell Signaling Technology); p70S6K (#9207, 1:1000, Cell Signaling Technology); Cdc45 (H-300 clone, SC20685, 1:50, Santa Cruz Technology); and phospho-Histone H3 (Ser10); (#9706, 1:100, Cell Signaling Technology).

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Qiu Z., Fa P., Liu T., Prasad C.B., Ma S., Hong Z., Chan E.R., Wang H., Li Z., He K., Wang Q.E., Williams T.M., Yan C., Sizemore S.T., Narla G, & Zhang J. (2020). A genome-wide pooled shRNA screen identifies PPP2R2A as a predictive biomarker for the response to ATR and CHK1 inhibitors. Cancer research, 80(16), 3305-3318.

Publication 2020

c-Myc (SC-40, anti-β-actin (Clone AC-74, anti-CHK1 (G-4, Histone H3 Phospho-eIF4E 4E-BP1 Phospho-4E-BP1 p70S6K phospho-histone H3 (Ser10);

4e bp1 Actin Antibodies Antibody Biological C myc Clone Eif4e Histone h3 P70s6k Phosphor Ppp2r5a Rabbit Rpa2 Western blot

Corresponding Organization : The Ohio State University

Other organizations : University of Rochester, Case Western Reserve University, University of Maryland, Baltimore, Augusta University Health, University of Michigan–Ann Arbor

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Protein expression levels of c-Myc, phospho-c-Myc (Ser62 and Thr58), RPA2, β-actin, CHK1, phospho-CHK1 (Ser345), CDC45, γ-H2Ax (Ser139), phospho-RPA32 (Ser4/Ser8), Histone H3, PPP2R5A, eIF4E, phospho-eIF4E (Ser209), 4E-BP1, phospho-4E-BP1 (Thr36/47), phospho-p70S6K (Thr389), p70S6K, phospho-histone H3 (Ser10)

control variables

None explicitly mentioned

positive controls

None explicitly mentioned

negative controls

None explicitly mentioned

Annotations

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