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Figure 6 | Retrovirology

Figure 6

From: Dephosphorylation of CDK9 by protein phosphatase 2A and protein phosphatase-1 in Tat-activated HIV-1 transcription

Figure 6

CDK9 is dephosphorylated by PP1 in cultured cells. A, Immunoprecipitation of CDK9. Lane 1, CDK9 was precipitated from HeLa cell extract with anti-cyclin T1 antibodies, resolved on 10% SDS-PAGE and immunoblotted with anti-CDK9 antibodies; Lane 2, immunoprecipitation of recombinant CDK9/cyclin T1; Lanes 3, input recombinant CDK9/cyclin T1; lane 4, input HeLa cell extract. B, HeLa cells were labelled with (32P) orthophosphate in the absence (lane 1) and in the presence of 1 μM okadaic acid (lane 2) and cellular extracts were immunoprecipitated with anti-cyclin T1 antibodies, resolved by 10% SDS-PAGE and transferred to PVDF membrane. Position of CDK9 was determined by probing the membrane with anti-CDK9 antibodies using 3,3'-Diaminobenzidine enhancer system. The picture is autoradiogram of the membrane exposed to phosphor imager screen. C, 293T cells were labeled with (32P) orthophosphate in the absence (lane 1) and in the presence of 100 nM okadaic acid (lane 2) and cellular extracts were immunoprecipitated with anti-CDK9 antibodies, resolved by 10% SDS-PAGE and transferred to PVDF membrane. Position of CDK9 was determined by probing the membrane with anti-CDK9 antibodies using 3,3'-Diaminobenzidine enhancer system. The picture is an autoradiogram of the membrane exposed to phosphor imager screen. D, 293T-cdNIPP1 cells stably expressing central domain of NIPP1 (143–224) were transfected with Flag-CDK9 expression vector and labeled with (32P) orthophosphate in the absence (lane 1) and in the presence of 100 nM okadaic acid (lane 2). Cellular extracts were immunoprecipitated with anti-Flag antibodies, resolved by 10% SDS-PAGE and transferred to PVDF membrane. Position of CDK9 was determined by probing the membrane with anti-CDK9 antibodies using 3,3'-Diaminobenzidine enhancer system. The picture is autoradiogram of the membrane exposed to phosphor imager screen.

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