Fig. 2

MiR-192-5p knockout human HCC cell lines presented the increased CSC features and were highly glycolytic. a Schematic structure of miR-192-5p wild-type (WT) and knockout (KO) by CRISPR/Cas9 system. The knockout of miR-192-5p DNA fragment was assessed by Sanger sequencing. Levels of miR-192-5p and miR-194 was measured by RT-qPCR. b Quantitative data of flow cytometry analysis using APC-conjugated antibodies against different CSC biomarkers in 192KO lines and WT lines from HLF and HLE cells. c Western blot of glycolytic enzymes using 192KO lines and WT lines from HLF and HLE cells. d ECAR and OCR were measured in 192KO lines and WT lines from HLF and HLE cells. e RT-qPCR analysis of miR-192-5p, flow cytometry analysis of CD24⁺ and CD44⁺ populations, and lactate measurement using Lactate assay kit in HLF cells with different levels of miR-192-5p. f Measurement of lactate in cultured medium from HLF and HLE cells at different time points. Representative images for color change of the corresponding cultured medium at 72 h after seeding are shown