Siglec-1 RNAi reduces VCC volumes in HIV-1 NLUdel-infected MDMs.
(A) MDMs were infected with VSV-G pseudotyped NLUdel. On the following day, MDMs were transfected with 60 nM scrambled, Siglec-1 or tetherin siRNA. At day 10 post-infection, MDMs were washed, fixed with 4% PFA and immunostained for p24 (green), Siglec-1 (red), tetherin (magenta) and DAPI co-stained. Size bars = 10 μm. (B) P24 VCC volume/cell from 30 NLUdel infected MDMs from each group were quantified using the Volocity 6.3 measurement module. Mean +/- SD shown for each RNAi group. (C) Scatter plot displaying individual p24 VCC volume measurements and mean from (B). (D) p24 release from Siglec-1, tetherin or control siRNA treated and NLUdel-infected MDMs was assessed over 12 days using a p24 ELISA. The efficiency of particle release is plotted as percentage of extracellular p24/total p24 from 3 experiments, with standard deviations indicated. Arrows indicate two timepoints when the media was changed, resulting in a transient drop in the ratio of extracellular p24/total p24.