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Figure 1 | BMC Molecular Biology

Figure 1

From: Analysis of chromatin boundary activity in Drosophila cells

Figure 1

Enhancer-mediated GFP activation is blocked by the suHw and the SF1 insulators in Drosophila S2 cells. A. Induction of S2 cells containing the GFP transgene. Top: a diagram of MT-GFP: key regulatory components shown include: the MT enhancer (yellow), the evenskipped basal promoter (light blue) and the GFP reporter gene (green). Middle: fluorescence microscopy images of S2 cell containing MT-GFP before (left) and after (right) induction. Bottom: FACS histogram of uninduced (left) and induced (right) S2 cells. X-axes: log scale of GFP level; Y-axes: cells number at indicated GFP level. Red bar: percentage of total cells with GFP level above 102. B. Induction of S2 containing MT-sp-GFP (grey box). Bottom: FACS histogram of uninduced (left) and induced (right) S2 cells. C-D. GFP induction in S2 cells containing enhancer-blocking transgenes. Top: a diagram of MT-suHw-GFP (C) or MT- SF1-GFP (D) transgenes. Insulator elements are represented by the red ovals. Bottom: FACS histogram of uninduced (left) and induced (right) S2 cells. E. Quantitation of GFP induction (I) in S2 cells transfected with insulator-containing transgenes. Number of replicates for each experiment is shown in parentheses. See methods for calculation of I and standard error of mean (SEM). F. Comparison of enhancer-blocking activity of suHw in S2 and Kc cells. Bar graph shows percentage of GFP induction in S2 (left) or Kc (right) cells transfected with MT-suHw-GFP (red bars) and MT-GFP (green bars). Transfection and induction were done in parallel.

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