Additional file 1: of Abi1 loss drives prostate tumorigenesis through activation of EMT and non-canonical WNT signaling

Figure S1. Evaluation of ABI1 expression in a test TMA. A) ABI1 expression is downregulated in prostate tumors with high Gleason grade pathology. The ABI1 expression levels were compared with the disease stage of the tumors. For KHSC cohort (n=32), binary scoring method was used as described in materials and methods section which categorizes patients into two (down/up) distinct groups based on arbitrarily defined threshold of % positive cells (>20%) and overall staining intensity as compared to internal positive controls (benign glands). Intra- and inter-core heterogeneity (i.e. mosaic pattern) were defined when cancer cells with decreased or retained ABI1 protein were observed within the same (intra) or different (inter) TMA cores and it was assessed manually. Subsequently, the case was considered homogeneous, when there was no inter-or intra-core heterogeneity seen. In this small TMA cohort within the cases with ABI1 down-regulation, we have observed 4 cases with inter-core heterogeneity, 2 cases with intra-core heterogeneity and one case with IDC with heterogeneous/mosaic pattern. (B) ABI1 expression and prostate tumor heterogeneity. Left, Contingency Table for ABI1 expression levels in Gleason patterns associated with different WHO grade groups. Gleason 3-3, Grade group 1; Gleason 3-4, Grade group 2; Gleason 3-3, Grade group 3; Gleason 4-4 and Gleason 5-3, Grade group 4; Gleason 4-5 and Gleason 5-5, Grade group 5. ABI1 expression was quantified by digital imaging as described in Material and methods and presented here as group of staining intensity: 0, negative staining, 1, weak staining; 2, moderate; 3, strong (digital scoring). Digital scoring takes into consideration % of cells stained for each level of intensity in a tumor core and is highly quantitative. The normalized stain intensity represents average intensity per core calculated as follow takes into consideration percent of cells stained for each intensity level (0, 1, 2 and 3), the average stain intensity for each core is calculated as sum of intensities and ranged from 0-1. The digital scoring showed statistically significant correlation with manual H-score as determined by direct comparison of digital vs. manual scoring of 505 patients in the TMA.The table below lists ABI1 expression level in tumors associated with each WHO tumor Grade Group histopathology, Group 1-5, and in benign tissue. On the right an image demonstrating intra-core tumor heterogeneity of ABI1 expression with Benign, an