Moreover, the apoptotic assay using Hoechst immunofluorescence staining led the same conclusion as the apoptotic assay using Annexin-V/PI staining for flow cytometric analysis, in which the HyPer-low subpopulation of CCA cells possesses a higher rate of gemcitabine chemoresistance (Fig. 2G, H). Here, ANXA5 is linked to cholangiocarcinoma.