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Fig. 2 | Retrovirology

Fig. 2

From: Effect of transcription inhibition and generation of suppressive viral non-coding RNAs

Fig. 2

Presence of ubiquitin-Tat in the large complex. a HIV-1 infected J1.1 cells were electroporated with CMV-Flag-Tat101 (20 μg) and kept at 37 °C for 48 h. Cells were isolated, washed, and extracts were processed for FPLC chromatography (Superose 6) using high salt. A total of 3.5 mg was used for chromatography. Flow rate parameters for the FPLC were set at 0.3 mL/min and 0.5 mL fractions of the flow-through were collected at 4 °C for approximately 60 fractions per sample (1 mL injected). Tat associated complexes were nanotrapped with NT084 and assayed for Western blot using α-Flag antibody. b Densitometry counts from panel a were obtained, normalized to background, and plotted to represent the relative abundance of Tat protein in each fraction. c Chromatography fractions were IPed with α-Flag antibody overnight, followed by addition of Protein A/G, ran on a gel, and analyzed by Western blot with α-ubiquitin antibody. Two sets of extracts (± F07#13) were run on chromatography and used for nanotrapping and Western blots

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