Table 1.

Immunohistochemical procedures used to detect Zeb1 and TIL subpopulations

Single labeling
Ag	Ab clone	Ab supplier	Ag retrieval	Ab dilution, incubation time, temperature of incubation	Detection system
Zeb1	EPR17375	Abcam	CC1 60 min	1/50, 60 min, 37°C	ultraView DAB
CD8	SP16	Thermo Scientific	CC1 36 min	1/200, 60 min, 37°C	ultraView DAB
CD20	SP32	CellMarque	CC1 36 min	1/100, 36 min, 37°C	ultraView DAB
FoxP3	SP97	Thermo Scientific	CC1 60 min	1/100, 60 min, 37°C	OptiView DAB
Double labeling
Ag	Ab clone	Ab supplier	Ag retrieval	Ab dilution, incubation time, temperature of incubation	Detection system
Zeb1	EPR17375	Abcam	CC1 60 min	1/50, 60 min, 37°C	UltraMap AR HRP + ChromoMap DAB
CD8	SP16	Thermo Scientific	CC1 36 min	1/200, 60 min, 37°C	UltraMap AR HRP + Purple Kit
CD20	SP32	CellMarque	CC1 36 min	1/100, 36 min, 37°C	UltraMap AR HRP + Purple Kit
FoxP3	SP97	Thermo Scientific	CC1 60 min	1/100, 60 min, 37°C	UltraMap AR HRP + Purple Kit

Legend: Ag, antigen; Ab, antibody; CC1, Cell Conditioning 1 (buffer); DAB, 3’, 3’-diaminobenzidine; HRP, horseradish peroxidase.