Figure 2.
Figure 2. S2 cell screen identifies genes that regulate the surface expression of dVMAT. (A) The flow chart illustrates the procedure used for screening. S2 cells transfected with GFP-/HA-dVMAT were treated twice with dsRNA over a 6-d period in a 96-well plate, incubated with external HA antibody conjugated to Alexa Fluor 647 for 2 h, washed, and the fluorescence of both GFP and Alexa Fluor 647 was measured at the level of individual cells by flow cytometry. Of 7,200 genes conserved from Drosophila to mammals, 18 positives (z score ≥ 3) were identified and retested using nonoverlapping dsRNA. Kolmogorov-Smirnov analysis of the cumulative frequency distribution reveals that 16 of 18 genes were again positive (P < 0.005 in at least two independent experiments). (B) Cumulative frequency distributions for selected dsRNA in representative retest experiments show the differences from control (wt) and DE/AA dVMAT.

S2 cell screen identifies genes that regulate the surface expression of dVMAT. (A) The flow chart illustrates the procedure used for screening. S2 cells transfected with GFP-/HA-dVMAT were treated twice with dsRNA over a 6-d period in a 96-well plate, incubated with external HA antibody conjugated to Alexa Fluor 647 for 2 h, washed, and the fluorescence of both GFP and Alexa Fluor 647 was measured at the level of individual cells by flow cytometry. Of 7,200 genes conserved from Drosophila to mammals, 18 positives (z score ≥ 3) were identified and retested using nonoverlapping dsRNA. Kolmogorov-Smirnov analysis of the cumulative frequency distribution reveals that 16 of 18 genes were again positive (P < 0.005 in at least two independent experiments). (B) Cumulative frequency distributions for selected dsRNA in representative retest experiments show the differences from control (wt) and DE/AA dVMAT.

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