Figure 3.

Expression of EAP45 in trans rescues the budding defect in HAP1 EAP45 KO cells. (a): EAP45 KO cells were transfected with WT provirus together with either 200 or 1,000 ng of HA‐tagged EAP45 expressor. Proteins from cell lysates or virions were detected by western blot using antibodies directly to HA, GAPDH, or p24. The viral products and the size markers are shown to the left of the blot. (b): The total cellular RNA was extracted followed by RT‐PCR for detection of the exogenously expressed EAP45 mRNA. Endogenously expressed GAPDH was also amplified and is shown as a loading control. (c): The release ratio is defined as the amount of p24‐associated products in purified virions divided by those in the cell lysate normalised to the level of GAPDH. The ratio is then normalised to that in the absence of EAP45. Error bars represent the standard error of the mean of four replicates from two independent experiments. Unless otherwise stated, statistical analysis was performed by using the two‐tailed student t‐test with significance annotated p ≤ .05 (*), p ≤ .01 (**), and p ≤ .001 (***) and ns (non‐significant) throughout this study. (d): The densitometric traces of p24 and p24‐p2 from (a) for each transfection condition are shown separately or in overlay to better visualise the differences in the final cleavage