Gliomas and related diseases biopsies were validated and obtained from Department of Pathology in Xiangya Hospital. IHC analysis of paraffin sections from gliomas tissues was described previously 38 (link). The sections were incubated with antibodies as indicated. The images were surveyed and captured using a CX41 microscope (OLYMPUS, Tokyo, Japan) with the Microscope Digital Camera System DP-72 (OLYMPUS, Tokyo, Japan) and differentially quantified by two pathologists who were from the Xiangya Hospital, Changsha, China.
GSK3β and LRP6 staining were considered positively by ascertaining cytoplasmic staining, whereas LSH and E2F1 were considered positively by nuclear expression. The determination results were obtained from semi-quantitative classification according to 10 or more visual fields (×200). The slides were first scored as 0 (negative), 1 (buff), 2 (pale brown), and 3 (tan). Positive expression of LSH, E2F1, GSK3β or LRP6 was scored as 0 (negative), 1+ (<10% of positively-staining tumor cells), 2+ (11-50% of positively-staining tumor cells), 3+ (50-75% of positively-staining tumor cells), and 4+ (>75% of positively-staining tumor cells). Both the scores by multiply were regarded as the determination result.
GSK3β and LRP6 staining were considered positively by ascertaining cytoplasmic staining, whereas LSH and E2F1 were considered positively by nuclear expression. The determination results were obtained from semi-quantitative classification according to 10 or more visual fields (×200). The slides were first scored as 0 (negative), 1 (buff), 2 (pale brown), and 3 (tan). Positive expression of LSH, E2F1, GSK3β or LRP6 was scored as 0 (negative), 1+ (<10% of positively-staining tumor cells), 2+ (11-50% of positively-staining tumor cells), 3+ (50-75% of positively-staining tumor cells), and 4+ (>75% of positively-staining tumor cells). Both the scores by multiply were regarded as the determination result.
