Fig. 3.
CD46 isomers rescue HCMV infection in ARPECD46-/- knockout cells. (a) Cell surface expression of WT ARPE-19, ARPECD46-/-, and ARPECD46-/- cells stably expressing CD46 C1, BC1, and BC2 (ARPECD46-/-C1, ARPECD46-/-BC1, and ARPECD46-/-BC2) was confirmed by fluorescence-based flow cytometry with anti-CD46 mAb GB24 followed by incubation with anti-mouse IgGAlexa647. The normalized cell number was plotted based on Alexa647 fluorescence intensity. (b) The cell lysates from the WT ARPE-19, ARPECD46-/-, ARPECD46-/-BC1, ARPECD46-/-BC2, and ARPECD46-/-C1 cells were subjected to anti-CD46 and GAPDH immunoblots, respectively. (c) HCMV AD169R and TB40/E infection of WT ARPE-19, ARPECD46-/-, ARPECD46-/-BC1, ARPECD46-/-BC2, and ARPECD46-/-C1 were assessed at 24hpi by Celigo cytometer. Percent infection was measured using an infectivity assay and WT ARPE-19 cells were used to normalize infection at 100 %. s.d. is depicted by errors bars. (d) A linear regression model was used to calculate the Goodness of Fit (R2 ) using infection (%) and mean fluorescence intensity (MFI) of CD46 surface expression of AD169R and TB40/E infected cells (c). *P<0.05, **P<0.01, ****P<0.0001 (two-way ANOVA, Sidak’s multiple comparison test) values are compared to ARPE-WT infection.