a, Illustration of the TMEM9 domains. b,
In silico analysis of TMEM9 expression in CRC. Oncomine analysis of TMEM9 expression in human cancers (www.oncomine.org). 10% gene rank; p-value<0.0001; fold change>2; compared with normal cells. c, High expression of TMEM9 in CRC. Two IECs and nine CRC cells were collected for IB and qRT-PCR. Tubulin served as a loading control, and TMEM9 expression was normalized by HPRT1. n=3 independent experiments.d, Expression of TMEM9 in mouse tissues. Protein and mRNAs were extracted from six mouse tissues and assessed by IB and qRT-PCR. CRC cells were served as a positive control. n=3 independent experiments e-g, Expression of TMEM29, CD44, and β-catenin in CRC TMA. IHC of TMEM9, CD44, and β-catenin was performed using TMA (Biomax, BC05023; e). H score (f) and Pearson’s correlations (g) were calculated. n=36 biologically independent samples.h, Co-expression of TMEM9 with AXIN2. Oncomine analysis of GSE20842, GSE5206, and TCGA data sets; 10% gene rank; P value<0.0001; fold change>2; compared with normal cells.i and j, Co-expression of TMEM9 with Wnt/β-catenin target genes. GEO data sets (GDS2947) were analyzed for each gene expression in normal intestine and the matched CRC samples (32 patient samples; i). Pearson’s correlations were determined (j).Scale bars=20μm; NS: Not significant. Error bars: mean ± S.D.; Two-sided unpaired t-test.