Table 2.

Quantitative analysis of SOX2 expression levels in areas with Patterns 1, 2 or 3 as determined by means of fluorescence intensity measurements (Arbitrary Units)

		Basal/Parabasal cell layers	Intermediate cell layers	Superficial cell layers
Pattern 1	Average	341*	11	14
	Range	75–499	3–26	3–35
Pattern 2	Average	1767	264	27
	Range	547–3202	85–496	4–89
Pattern 3	Average	51	498	54
	Range	2–108	62–878	3–143

Fluorescence intensity was measured after an immunohistochemical staining procedure using FITC-tyramide as a peroxidase substrate. The SOX2 expression intensity was measured in 5 different sections. Each cell layer in a total of 50 nuclei was measured. Nuclei were not overlapping, nuclear truncation was minimal (large nuclear size) and on average 3.5 slices per nucleus (3–5 slices) were used for reconstruction. In total, 450 nuclei were reconstructed and measured (*50 nuclei per SOX2 pattern and per cell compartment). For quantification, the images are captured with the same fluorescence integration time during confocal microscopic imaging. The average fluorescence intensity and the range of fluorescence intensity are expressed in arbitrary units. ImageJ (NIH, Bethesda, Maryland, USA) was used for further image analysis, processing and merging/stitching of the fluorescent images for reconstruction of the sections