Fig 1: Elevated GANAB expression was significantly correlated with poor prognosis of UCs. A, Representative images to indicate the categorical IHC staining intensities. Score 0: Negative; Score 1: Weak staining; Score 2: Medium staining; Score 3: Strong staining. Scale bar: 100um; magnification at 200 × (top). Scale bar: 25µm; magnification at 800 × (bottom). B, The higher expression levels of GANAB was significantly correlated with poorer outcome of UCs in our cohort. Note, the patients with score 0 were grouped into the negative expression group; the patients with score 1–3 were grouped into the positive expression group. C-D, The high H-score group was significantly associated with the poorer survival of UCs in our cohort. The median cut-off value (C) and the optimal cut-off value (D) were used, respectively. E, Multivariate analysis of the Glycoprotein geneset using the Cox regression model based on filtered variables. F, Survival curves between high risk and low risk groups of the UC patients based on the risk scores in the Glycoprotein-based mRNA model. Log-rank test was used for the significant test. G, Survival curves between the high-expression and low-expression groups, stratified by the gene expression levels of GANAB in the TCGA-BLCA cohort. The optimal cut-off value of GANAB mRNA expression was used
Fig 2: High histological grade and MIBC were correlated with elevated GANAB expression levels in UCs. A-B, The UCs of low-grade showed lower expression levels of GANAB, as compared with the UCs of high-grade. C-D, MIBC showed higher expression levels of GANAB as compared with NMIBC. IHC Scale bar: 25µm; IHC magnification at 800 × . HE Scale bar: 500µm; HE magnification at 50 × . IHC assay IDs were indicated in yellow. NMIBC = Non-muscle invasive bladder cancer; MIBC = Muscle-invasive bladder cancer. *p < 0.05; ****p < 0.0001
Fig 3: Knockdown of GANAB induced ER stress and dys-regulation of cell cycle genes in vitro. A, The expression of GANAB was up-regulated in T24 and UM-UC-3 cells with tunicamycin (Tm) stimulation. B, The expression of HIF1A in both siGANAB and Tm treatment was higher than that in cells with Tm treatment alone. C, The expression of ATF6 in both siGANAB and Tm treatment was higher than that in cells with Tm treatment alone. D, The expression of E2F7 was up-regulated in cells with siGANAB or Tm treatment. E, The expression of FOXM1 was down-regulated in siGANAB or Tm treatment. *p < 0.05, **p < 0.01, ***p < 0.001
Fig 4: GANAB promoted the proliferation in UC cells. A-B, GANAB was markedly down-regulated or up-regulated after siGANABs or pcDNA3.1( +)-GANAB transfection. C-D, SiRNA knockdown of GANAB in T24 and UM-UC-3 cells significantly inhibited cell proliferation in CCK8 assay. E–F, Overexpression of GANAB in T24 and UM-UC-3 cells markedly increased cell proliferation in CCK8 assay. G-H, Colony-formation assays demonstrated that knockdown of GANAB dramatically inhibited the size and the number of colonies of T24 and UM-UC-3 cells. I- J, The overexpression of GANAB increased the size and the number of colonies of T24 and UM-UC-3 cells. *p < 0.05; ***p < 0.001
Fig 5: Bioinformatics analysis of the expression regulatory factors of GANAB. A-B, GANAB expression was up-regulated by the copy number amplifications of the gene loci of the UC genomes. C-D, The expression of the GANAB was positively correlated with the gene signature-derived Hypoxia scores in TCGA-BLCA data (CBioportal). E–F, The expression levels of the GANAB was positively correlated with Stress Granule (SG) factors (G3BP1 and GEMIN5). The color scheme used in C-F was the same as B. ‘-’, not profiled, or no mutation detected in the genomic data
Supplier Page from Abcam for Anti-GANAB antibody [EPR12376]