Table 2 Unpaired t test for mean vista expression. Significant features (P ≤ 0.05) are shown in bold.

Variable	VISTA manual negative	VISTA manual positive	P-value	VISTA superpixel negative	VISTA superpixel positive	P-value
VISTA expression
Age			0.086			0.817
≤ 70	97	35		89	43
> 70	59	35		62	32
Gender			0.100			0.167
Male	75	43		73	45
Female	81	27		78	30
Pre-surgery chemotherapy			0.880			0.899
No therapy	146	67		142	71
Complete regression	0	0		0	0
Partial regression	7	2		6	3
No regression	3	1		3	1
Pre-cancer condition			0.918			0.938
Non	94	44		92	46
TA	1	1		1	1
VA	46	19		44	21
TVA	14	6		13	7
SSA	1	0		1	0
AJCC stage			0.022			0.001
I/II	71	42		67	46
III/IV	85	28		84	29
Pathologic stage			0.099			0.052
PT1	12	5		13	4
PT2	14	4		15	3
PT3	83	48		79	52
PT4	47	13		44	16
Lymph node stage			0.719			0.753
N0	96	44		92	48
N1–2	60	26		59	27
Tumor grade			0.232			0.292
G1–G2	115	45		111	49
G3	41	25		40	26
Tumor budding			0.670			0.757
TBD1	92	44		87	49
TBD2 or TBD3	64	26		64	26
LVI			0.498			0.541
Not present	90	37		87	40
Present	66	33		64	35
TIL			0.001			0.001
Low	97	15		96	16
High	59	55		55	59
Stromal differentiation			0.007			0.001
Immature	86	25		87	24
Mature	70	45		64	51
MMR status			0.003			0.003
Intact	137	50		133	54
Loss	19	20		18	21
KRAS			0.867			0.667
Wild type	14	5		14	5
Mutation	11	4		11	4
BRAF			0.001			0.031
Wild type	15	1		14	2
Mutation	5	11		7	9

VISTA V-domain immunoglobulin suppressor of T cell activation, TBD tumor budding grade, PT stage, N nodal stage, LVI lymph-vascular invasion, TIL tumor-infiltrating lymphocytes, G grade, t test for selected biomarkers, MMR mismatch repair gene, KRAS Ki-ras2 Kirsten rat sarcoma viral oncogene homolog, BRAF B-Raf Proto-Oncogene.

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