Figure 7

BRG1 and Tat associate with chromatinized HIV-1 promoter in C33A cells at nuc-1. A) The BRG1/hBrm-deficient cervical carcinoma cell line C33A was used for these experiments and maintained under standard conditions in Dulbecco's modified Eagle's medium. A BRG1 (10 μg) or Tat (3 μg) expression vector or a control vector (10 μg, lane 3) was transfected into C33A cells that have an HIV-1 reporter construct stably integrated into their genome. After cells were allowed to express BRG1 for 36 h, cultures were stimulated with either PMA for 2 h (lane 5), TSA for 12 h (lane 6), or both for 2 h and 12 h, respectively (lane 7), before ChIP. A ChIP assay was performed with BRG1 antibodies, and PCR was used to detect the recovery of nuc-0 and nuc-1 DNA. The input DNA represents the total genomic DNA. Ab, antibody; Unstim, unstimulated. B) BRG1 and Tat were transiently expressed in C33A cells that had been stably transfected with an HIV-1-luciferase construct. Thirty-six hours after transfection, cells were stimulated with either PMA for 2 h, TSA for 12 h, or both for 12 h. Following stimulation, cells were lysed, and the luciferase activity was measured [114]. Data presented is from average of two individual experiments.