TABLE 2.

Regulation of B7-H1 expression in vestibular schwannoma tumors: real-time polymerase chain reaction cycle-thresholds (Ct) for 5 strongly positive (2+) immunohistochemistry-stained B7-H1 tumors compared with 5 B7-H1 absent (0) vestibular schwannoma specimens

Vestibular schwannoma specimen	miR-513 Ct	U6 Ct	miR-513 Ct/U6 Ct	B7-H1 mRNA Ct	B7-H1 copy/µl	Actin copy/µl	B7-H1/actin ratio
B7-H1 (2+) A	59.6	29.88	1.99	35.0	1440	4300	0.33
B7-H1 (2+) B	46.7	32.11	1.45	35.1	1380	1790	0.77
B7-H1 (2+) C	43.31	31.71	1.37	37.2	716	751	0.95
B7-H1 (2+) D	ND	ND	ND	34.8	1580	3200	0.49
B7-H1 (2+) E	ND	ND	ND	33.1	4060	4250	0.96
Average (2+)			1.60 ± 0.34 (p = 0.62)				0.70 ± 0.28 (p = 0.35)
B7-H1 (0) F	46.84	33.13	1.41	35.9	1020	1140	0.89
B7-H1 (0) G	49.47	30.29	1.63	35.9	1020	2310	0.44
B7-H1 (0) H	41.51	30.31	1.37	36.3	900	880	1.02
B7-H1 (0) I	ND	ND	ND	34.7	1690	1130	1.50
B7-H1 (0) J	ND	ND	ND	36.3	900	1130	0.80
Average (0)			1.47 ± (p = 0.62)				0.93 ± 0.38 (p = 0.35)
H69	47.15	27.84	1.69	ND	ND	ND	ND
Tonsil	0	30.21	0	32.1	2750	13200	0.21

RNA was available for only 3 positive and negative tumors for miR-513, which were normalized with small nuclear RNA RNU6 (U6). All 10 tumors were used to evaluate B7-H1 mRNA expression. Ct values were normalized to a standard curve of serial dilutions of a known B7-H1 RNA concentration (copy/µl). This was then normalized against actin mRNA copy/µl in the last column. Human cholangiocyte cells (H69) were used as a positive control, and human tonsil tissue was used as a negative control. No difference in the quantity of miR-513 or B7-H1 mRNA was detected between groups of strongly positive and negative B7-H1 protein immunostained VS (p = 0.62 and p = 0.35, respectively).

Bold values are the average miR-513 Ct/U6 Ct and B7-H1/actin ratio values for the B7-H1 2+ and negative (0) immunostaining schwannoma tumors.

ND indicates not determined.