TABLE 2.
Comparison of Gag protein production and cytoplasmic accumulation of HIV RNAa
| Plasmid
|
PhosphorImager units (105) normalized to gapdh RNA signal
|
||||||
|---|---|---|---|---|---|---|---|
| Total
|
Cytoplasmic
|
Foldc
|
|||||
| Name | Gagb (pg) | Unspliced | Spliced | Unspliced | Spliced | Unspliced | Spliced |
| pYW99 | <MDd | 16.6 (1.0e) | 51.3 (3.1) | 2.3 (1.0) | 14.1 (6.1) | 1.0 | 1.0 |
| pYW208 | 8,000 | 6.4 (0.4) | 23.0 (1.4) | 3.0 (1.3) | 23.4 (10.2) | 3.3 | 4.3 |
| pYW205 | 12,000 | 10.4 (0.6) | 43.8 (2.6) | 1.8 (0.8) | 14.6 (6.3) | 1.3 | 1.1 |
| pYW100 | 81,000 | 9.1 (0.5) | 56.0 (3.4) | 2.4 (1.0) | 35.6 (15.5) | 2.0 | 2.3 |
a
Total cellular or cytoplasmic RNAs were harvested from duplicate cultures of 106 293 cells at 2 days posttransfection and subjected to DNase treatment. Fifteen-microgram aliquots were subjected to RPA with uniformly labeled antisense HIV and gapdh RNA probes, PAGE, and PhosphorImager analysis.
b
Cell-associated Gag protein measured by Gag ELISA.
c
Cytoplasmic RNA/total RNA level relative to cytoplasmic RNA/total RNA level of pYW99.
d
<MD, less than the minimum detectable.
e
RNA level presented relative to level of pYW99 unspliced RNA.