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In addition, the diffraction pattern shows no twin spots, confirming a lack of twin defects.
The inserted atomic simulation of two slabs 2 nm in thickness superimposed by 52.2 degree off ind indeed shows lamellae-like fringes with corresponding twin spots (denoted as t) in the simulated diffraction pattern ine.
Nevertheless, the presence of twin spots (marked by high levels of GFP) (Figure 1D', encircled) indicated that mutant clones were induced, but not recovered.
Clone sizes were quantified based on the number of cells/nuclei per clone (observed by DAPI staining) and average cell number per clone and standard deviations were graphed using Microsoft Excel for both clones and twin spots.
We also identified a number of cell lethal mutants that generated wild type twin spots but no apparent corresponding mutant clone, implying that recombination had occurred but that the mutant cells did not survive.
To verify that the low rate of recovered clones in the MARCM experiments was not due to the experimental conditions control clones were induced in parallel using FRT40A ubi-GFP to monitor the appearance of twin spots in the wing disc.
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P-values for twin spot analysis were obtained using a two tailed paired t-test.
Mutant clones and twin spot areas were measured with confocal images using the histogram function of Adobe Photoshop.
Cell proliferation was assessed by comparing the number of Slbp15 mutant to the total number of cells in each twin spot.
To test whether Slbp mutants exhibit a proliferation defect, we performed a "twin spot" analysis in wing imaginal discs by inducing mitotic clones using FLP recombinase [52].
Mutant clones were identified by the lack of GFP and the close proximity of twin spot clones with two copies of the GFP expression cassette.
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