FIG 9.

Abnormalities caused by CRE-kB and add-on RARE mutations also manifest in D-NT2. (A) HFF (MOI, 0.7 PFU/cell) and D-NT2 (MOI, 1.0) were infected with rWT, rCK⁻, and rCKR⁻ in the absence of R exposure. FACS was applied at day 1 p.i. to determine the percentages of MIE⁺ cells. Scatter plots show percentages of 250,000 gated cells in bright and dull MIE^{+ −}D-NT2 subsets, which make up the total MIE⁺ population (MIE⁺ Tot.). The bar graph depicts the ratio of bright to dull MIE⁺ D-NT2. The histogram shows the fluorescence intensity profile for the MIE^Bright D-NT2 population. (B) At 1 day p.i. with rCK⁻.A, rCK⁻.B, rCKR⁻.A, and rCKR⁻.B, spliced MIE RNA levels were quantified from triplicate infections of HFF (MOI, 1.0) and D-NT2 (MOI, 0.03) in the absence of R and normalized to actin RNA levels. (C) D-NT2 (MOI, 0.03) infected with rWT, rCK⁻, and rCKR⁻ in parallel with groups shown in panel A were maintained in the presence of R for 14 days p.i. The average number (Avg. No.) of vGFP⁺ D-NT2 clusters (>6 vGFP⁺ D-NT2 per cluster) was determined for two separate infections. Size distributions of 50 randomly selected vGFP⁺ NT2 clusters are depicted by category of diameter size. rCKR⁻ did not form D-NT2 clusters at day 14. (D) HFF (MOI, 0.6 PFU/cell) and D-NT2 (MOI, 0.3 and 0.03) were infected with rWT, rC⁻, rK⁻, and rCK⁻. FACS determined the percentage of MIE⁺ D-NT2 at 1 day p.i. The average numbers of vGFP⁺ D-NT2 clusters were determined at day 14 p.i. for two separate infections.