VHH05 blocks phosphorylation of UBC6e when expressed in mammalian cells. (a) Phosphorylation of UBC6e in HeLa was reduced by cytosolic expression of VHH05, but not VHH68 that is specific to the nuclear protein of influenza virus. Treatments: 1% DMSO for 30 mins, 50 μM cycloheximide (CHX) for 30 mins, 10 mM DTT for 1 hour, 10 μM Thapsigargin (Tg) for 2 hours, 25 μM MN2743 (E1I, E1 inhibitor) for 1 hour. (b) VHH05 slows and dampens the phosphorylation of UBC6e when HeLa cells were treated with 50 μM CHX for the time indicated. Quantified data (right) depict mean ± standard deviation and lines serve only to guide the eye. *p<0.05 for 6E05 vs VHH68; **p<0.005 for 6E05 vs VHH68 at the indicated time points.