FIG. 1.

Dose, time and mucins effects on primary HAE BAAV TA mediated transduction. (A and B) Monolayer of differentiated HAE, plated in 6mm transwell filters, were incubated apically with 10⁸ DNAse resistant particles (DRP) of BAAV-GFP and treated on the basolateral surface with 0, 0.031, 0.062 or 0.125, % w/v of TA for 3 hrs. 48 hrs post incubation, GFP positive cells were observed by fluorescent microscopy and fluorescence quantified using the ImageJ software. (C and D) As above, cells were incubated apically with BAAV-GFP but treated with 0.015% w/v of TA for 3 or 24 h respectively. 96 hrs later GFP positive cells were observed and quantified. (E, F and G) HAE cultures secreting mucins were extensively washed with cell medium or left untreated. BAAV was applied apically and cell treated on the basolateral surface with 0.25 or 0.5% w/v TA for 3 hrs. 48 (E) and 96 (F) hrs later positive cell were observed and quantified. N=2 in duplicate. Student’s t-test * (P value <0.05). Positive cells of 4 random fields in experiment A were also counted and similar fold changes compared with control were measured (data not shown).