Figure 3.
Figure 3. No evidence for extended looping of BAC sequences from a linear backbone. (A–C) 3D DNA FISH shows co-linear signals from the Lac operator (red) and BAC (green) probes. DNA is stained by DAPI (gray). (A) DHFR D10 cells. (B) DHFR-110-5-E4 cell with lac operator repeats inserted into the DHFR gene fifth intron. (C) Hsp70_5 cell 30 min after heat shock. Middle columns (deconvolved data) and right columns (raw data) show 2x enlarged views (Bars, 2 µm). (D) EM of DHFR D10 clone by in vivo immunogold staining of EGFP-LacI shows the BAC transgenes are compacted indistinguishable from surrounding genomic DNA, with no obvious looping structure from the transgene backbone. Left and middle: serial sections (150 nm); right: 500-nm section (Bar, 500 nm).

No evidence for extended looping of BAC sequences from a linear backbone. (A–C) 3D DNA FISH shows co-linear signals from the Lac operator (red) and BAC (green) probes. DNA is stained by DAPI (gray). (A) DHFR D10 cells. (B) DHFR-110-5-E4 cell with lac operator repeats inserted into the DHFR gene fifth intron. (C) Hsp70_5 cell 30 min after heat shock. Middle columns (deconvolved data) and right columns (raw data) show 2x enlarged views (Bars, 2 µm). (D) EM of DHFR D10 clone by in vivo immunogold staining of EGFP-LacI shows the BAC transgenes are compacted indistinguishable from surrounding genomic DNA, with no obvious looping structure from the transgene backbone. Left and middle: serial sections (150 nm); right: 500-nm section (Bar, 500 nm).

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