Figure 3.

A) Generation of HOS_A4 cells by transduction with HIV-Intro and selection as described in the text. Tat induction induced the expression of ECFPskl in the cytoplasm. Top panels: phase contrast. Bottom panels: ECFP channel. B) Southern blot analysis of HOS_A4 cells shows the presence of a single integration event. Genomic DNA was digested with XhoI or SpeI and hybridized with a probe encompassing ECFP. C) Effect of Tat-mDsRed on HOS_A4 cells. Co-localization of Tat and HIV-Intro RNA is shown on the single transcription spot present in HOS_A4 cells. Correct gene expression is demonstrated by the ECPFskl signal in the cytoplasm. D) Co-localization of RNAPII and Cyclin T1 on HOS_A4 transcription spots. Cells were transfected with pEYFP-MS2nls and Tat, fixed and Cyclin T1 (top panels) or RNAPII (bottom panels) detected by immunofluorescence as described in [30].