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. 2018 Feb 23;8:3521. doi: 10.1038/s41598-018-21942-5

Figure 5.

Figure 5

BI-2536 and BI-6727 activate HIV-1 LTR transcription probably through bromodomain inhibition rather than PLK inhibition. (a) #95 and #225 THP-1-NanoLuc clones were cultured in 0.1% dimethyl sulfoxide (DMSO) in the absence or presence of NMS-P937 or PLKi-III at 0.01, 0.03, 0.1, 0.3, 1, 3, and 10 μM for 24 hours. The clones were also cultured with 1 μM BI-2536 or 1 μM BI-6727. After the clone cells were harvested and lysed, the NanoLuc activity of cell lysates was determined. Each value is shown as a mean ± standard deviation (SD) of triplicate samples. (b) #95 and #225 THP-1-NanoLuc clones were cultured without (−) or with (+) 1 μM BI-2536 or 1 μM BI-6727 in the absence (−) or presence (+) of 1 μM JQ1 or 10 μM SAHA for 24 hours. After the cells were harvested and lysed, NanoLuc activity of the cell lysates was determined. Each value is shown as a mean ± SD of triplicate samples. P-values were determined using Student’s t-test. P-values are significant if less than 0.05. (c) #95 THP-1-NanoLuc clone cells were treated with 0.1% DMSO, 1 μM BI-2536 or 1 μM BI-6727 for 24 hours and subjected to ChIP assay. The level of BRD4 protein associated with the HIV-1 LTR promoter was determined. ChIP DNA signals were normalized by those of input DNA. Each value is shown as a mean ± SD of triplicate samples. P-values were determined using Student’s t-test. P-values are significant if less than 0.05.