Mutant
|
No. of cell profiles analyseda
|
Cell surface
|
IPMC
|
% of buds in IPMC
|
Enrichment of buds in IPMCb
|
---|
No. of virus buds
|
μm of membrane
|
Buds/μm of membrane
|
No. of virus buds
|
μm of membrane
|
Buds/μm of membrane
|
---|
PTAP–
|
D415c
|
7 (5)
|
26
|
2620
|
0.0099
|
1137
|
1378
|
0.8248
|
97.8
|
83 ×
|
PTAP–YP–
|
D428
|
9 (5)
|
4
|
2041
|
0.0020
|
719
|
931
|
0.7723
|
99.4
|
394 ×
|
D429
|
12 (11)
|
89
|
5278
|
0.0169
|
1149
|
1814
|
0.6333
|
92.8
|
38 ×
|
D432
|
6 (5)
|
44
|
2486
|
0.0177
|
417
|
483
|
0.8633
|
90.5
|
49 ×
|
Overall total
|
34 (26)
|
163
|
12,424
|
0.0131
|
3422
|
4607
|
0.7428
|
95.5
|
57 ×
|
- Monocyte-derived macrophages (MDMs) from four different donors were infected with the HIV-1 R3A mutants PTAP– or PTAP–YP–, and infected MDMs located by on-grid immunolabelling. Electron microscopy images were recorded and analysed by counting the number of virus buds seen either at the cell surface or within intracellular plasma membrane-connected compartment (IPMCs). Linear lengths of surface or IPMC membrane within the sections were measured by tracing (see Methods)
-
a The number of cell profiles for which viral buds were counted and membrane lengths measured (the number of distinct cells analysed is given in brackets)
-
b The ratio of the density of buds/μm of membrane in the IPMC compared to the cell surface is shown as fold enrichment
-
c Donor ID