Immunoblotting of Cell Cycle Regulators

Immunoblotting was carried out as previously described [40 (link)] using the following primary antibodies: CDK7 (Cell Signaling Technology, 2916), CDK1 (Cell Signaling Technology, 9116), CDK2 (Cell Signaling Technology, 2546), CDK4 (Cell Signaling Technology, 12790), CDK9 (Cell Signaling Technology, 2316), β-actin (Abcam, ab6276), GAPDH (Cell Signaling Technology, 2118), P-S5 PolII (Abcam, ab5401), P-S2 PolII (Abcam, ab5095), PolII (Abcam, ab817), P-S780 Rb (Abcam, ab47763), P-S807/811 Rb (Cell Signaling Technology, 8516), Rb (Abcam, ab6075), cyclin H (Abcam, ab54903), MAT1 (Santa Cruz Biotechnology, sc135981), P-S15 p53 (Cell Signaling Technology, 9284), p53 (Santa Cruz Biotechnology, sc-126), P-T161/T160 CDK1/CDK2 (Abcam, ab201008), P-S10 Histone H3 (Abcam, ab139417), AR (MilliporeSigma, 06-680), P-T1457 MED1 (Abcam, ab60950), MED1 (Bethyl Laboratories, A300-793A), cleaved PARP1 (Abcam, ab4830), p21 (Santa Cruz Biotechnology, sc-817).

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Constantin T.A., Varela-Carver A., Greenland K.K., de Almeida G.S., Olden E., Penfold L., Ang S., Ormrod A., Leach D.A., Lai C.F., Ainscow E.K., Bahl A.K., Carling D., Fuchter M.J., Ali S, & Bevan C.L. (2023). The CDK7 inhibitor CT7001 (Samuraciclib) targets proliferation pathways to inhibit advanced prostate cancer. British Journal of Cancer, 128(12), 2326-2337.

Publication 2023

β-actin GAPDH P-S5 PolII P-S2 PolII PolII P-S780 Rb P-S15 p53 P-T161/T160 CDK1/CDK2 cleaved PARP1

Actin Antibodies Cdk1 Cdk2 Cdk9 Cyclin h Gapdh Histone h3 Med1 Parp1 T160

Corresponding Organization :

Other organizations : Hammersmith Hospital, Imperial College London, MRC London Institute of Medical Sciences

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Variable analysis

independent variables

Primary antibodies used for immunoblotting:

dependent variables

Protein expression levels of CDK7, CDK1, CDK2, CDK4, CDK9, β-actin, GAPDH, P-S5 PolII, P-S2 PolII, PolII, P-S780 Rb, P-S807/811 Rb, Rb, cyclin H, MAT1, P-S15 p53, p53, P-T161/T160 CDK1/CDK2, P-S10 Histone H3, AR, P-T1457 MED1, MED1, cleaved PARP1, p21

control variables

Not explicitly mentioned

controls

No specific positive or negative controls were mentioned in the provided information.

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