Fig 1: Expression of carbonic anhydrase 2 (CA2) in the urinary bladder of Hras 128 rats. Normal epithelial cells in the control rats were negative for CA2 (a). CA2 staining was localized to the cytoplasm of urothelial carcinoma (UC) induced by BBN→PEITC treatment ((b) HE staining; (c) CA2 staining). Summary of the expression of CA2 in rat UC induced by BBN→PEITC treatment (d).
Fig 2: Statistical modeling identifies signatures of differentially expressed proteins.A, two-pronged modeling identifies 13 proteins that are significantly differentially expressed between patients with DENV and ZIKV infection (adjusted p-value = 0.05). The heatmap shows the row-wise clustered and scaled residual abundance data after removing confounding effects for the 47 samples with complete meta-data. The extended abundance data for the 13 proteins is shown in supplemental Table S3. B, western blotting confirms differential expression for Carbonic anhydrase II (CA2) for patient samples from both the original cohort (2016/2017) used for the proteomics analysis and a new, independent cohort (2017/2018). Not shown: one extreme value for Zika (2017/2018 cohort) with normalized expression of 6.82. Anti-CA2 western blot signals were normalized by signal of reference Ponceau staining. Crosses within the box-and-whiskers plots indicate the average. p-values were calculated for two-tailed t test of normalized expression values of dengue versus Zika samples within a cohort. Full images are shown in supplemental Fig. S8. Quantitation is provided in supplemental Table S7. C, using the 13 differentially expressed proteins and a subset of three most predictive proteins, we evaluated model performance on an independent test set. For the classification with three most predictive proteins, we obtained 70% true-positive Zika predictions at a 6% false-positive rate (indicated by red dashed line). The area underneath the receiver–operator curve is 0.81. D, correctly predicted samples with DENV and ZIKV infections differ in the intensity (log2 transformed) of the three best proteins. Mispredicted samples have intermediate intensities. E, our predictions are confirmed by outside evidence. Additional experimental testing on a subset of samples (supplemental Table S6) revealed cases of ambiguous diagnosis: samples with equivocal results, samples of dengue diagnosis, which might be Zika infections or which may also have signs of past infections, and some Zika samples with an indication for other febrile infections. The graph evaluates our predictions with respect to the original diagnosis and these additional findings. Supplemental Fig. S5 shows this graph for the test set only.
Fig 3: Expression of carbonic anhydrase 2 (CA2) in the human urothelial carcinoma (UC). Normal epithelial cells were negative for CA2 (a). CA2 staining was localized to the cytoplasm of UC ((b) HE staining; (c) CA2 staining). Analysis of the cumulative incidence of stage progression of 95 non-muscle-invasive bladder cancer (NMIBC) patients after transurethral resection of bladder tumor (TURBT) showed that CA2-positive UC had a more rapid disease progression than CA2-negative UC: progression being defined as an increase in stage of pTa to =pT1 or pT1 to =pT2, (P < 0.0001, HR = 10) (d).
Supplier Page from Abcam for Anti-Carbonic anhydrase 2/CA2 antibody