Fig. 3

The expression of ASS1 protein in HLSCs and ASS1-HLSCs. a Western blot confirming the presence or absence of ASS1 protein isoform 1 (Iso1) and isoform 2 (Iso2) in 10 μg total protein extracted from human hepatocytes (hH), human liver stem cells (HLSCs), HLSCs from the liver of a patient suffering from citrullinemia type 1 (ASS1-HLSCs), and Rotary Cell Culture System (RCCS)-differentiated HLSCs and ASS1-HLSCs. Actin was run on a parallel gel as a representative of loading. b Quantification of the relative band intensity of ASS1 Iso1 across all samples in Fig. 3a normalized to actin. c Representative immunofluorescence micrographs of hH, HLSCs, ASS1-HLSCs, and RCCS-differentiated HLSCs and ASS1-HLSCs stained for ASS1 enzyme. The nuclei of the cells were stained with Hoechst dye 33258. Original magnification × 400, scale bar = 50 μm. Data represent one of two experiments with similar results. d Quantification of the relative fluorescence intensity (RFI) of all the cell culture conditions in Fig. 3c with hH serving as a positive control established as the 100%. Data represent the mean of three independent experiments with similar results. ^§ p < 0.05, HLSCs/RCCS vs HLSC; ^{^} p < 0.05, ASS1-HLSC/RCCS vs ASS1-HLSC; *p < 0.05, HLSC/RCCS vs HLSC; ^# p < 0.05, ASS1-HLSC/RCCS vs ASS1-HLSC