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. 2001 Jun 1;15(11):1383–1392. doi: 10.1101/gad.901101

Figure 4.

Figure 4

TSC1− and TSC2− cells are diploid. In A–D, mutant clones were generated by FRT/FLP and marked by the absence of Ubi-GFP signal (green). (A–B) Confocal images of a portion of a third-instar eye disc containing two TSC1− clones (arrows). The disc was stained with DNA dye propidium iodide (PI, shown in red). Two images are shown, one of GFP (A) and one of PI staining (B). Note that the intensity of PI staining is weaker in the mutant cells. Quantitation of Z series by using LSM 510 software showed that the total fluorescence in the nuclei is comparable between the mutant and the wild-type cell (data not shown). (C–D) Fluorescent light microscopy images of a portion of a third-instar eye disc containing a TSC2 (gigas) mutant clone (arrow). The disc was stained with DNA dye Hoechst 33342 (blue). Note that the intensity of Hoechst 33342 staining is weaker in the mutant cells. Quantitation by using the Axiovision software showed that the total fluorescence in the nuclei is comparable between the mutant and the wild-type cell (data not shown). (E–F) Flow cytometric analysis of dissociated wing imaginal discs containing TSC1− (E) and TSC2− (F) clones. The profiles of mutant and wild-type (WT) cells are indicated by heavy and light traces, respectively. No signal was detected beyond the DNA content of cycling diploid cells (data not shown). Note that the DNA content of TSC1− or TSC2− cells is similar to that of wild-type cells.