Table 2.

Data analysis and normalization: (A) Technical and biological sources of variation during the digital spatial profiling (DSP) working process. To subtract the variation, NanoString Technologies offers ‘different types’ of normalization for protein DSP; (B) Isotype controls and housekeeping proteins included in the DSP probe panel

A
	Source of variation	Type of normalization
Technical variation	nCounter quantification	•ERCC correction
Biological variation	ROI size	•Area normalization
	ROI cellularity	•Reference protein/gene normalization •Nuclei count normalization
	ROI background	•Three isotype controls

B
Isotype controls	Mouse IgG1, IgG2a and rabbit IgG
Housekeeping proteins	Histone H3, GAPDH and ribosomal protein S6

ROI, region of interest; ERCC, External RNA Control Consortium.