Figure 6. FIX-ΔLUNA viral infection of CD14+ cells fail to produce infectious virus after IL6 differentiation.
A) Cell lysates plus supernatant from FIX-WT, FIX-ΔLUNA, or FIX-Rev infected cells were collected at 10 dpi and 20 dpi. IL6 was added at 10 dpi; at 5 (15dpIL6) and 10 (20dpIL6) days after differentiation, cell lysates and sups were also collected. The collected lysate/supernatant mix was serially diluted and used to infect HF cells. Two days post infection, cells were fixed with 4× paraformaldehyde, then stained with mouse anti-IE1/2 (1∶250) overnight. Cells were then incubated with Alexa Fluor 488 anti-mouse IgG for 1 hr, and incubated with DAPI for 30 min. IE positive cells were counted to determine the number of infectious units (IU). Only the time points post reactivation showed any significance when comparing FIX-WT or FIX-Rev to FIX-ΔLUNA (10dpIL6: p<3×10−5; 20dpIL6: p<0.0006). The ** indicates a P-value<0.001.