Fig 2: The HFn–CTX nanoconjugate and the HFn nanoparticles show similar uptake kinetics in GBM cells. (A) Representative images of U-87 MG cells incubated with double-labeled HFn–CTX or labeled HFn (0.1 mg mL–1 each) for 2, 5, 25, 30, 60, and 120 min. HFn–AF647 is depicted in red, CTX in magenta, and CTX detected with anti-human-AF488 in green. The plasma membrane (PM) was stained with CD44 (yellow), and nuclei were stained with Hoechst (blue). Scale bar, 100 μm. (B) Manders’ coefficient analysis on the HFn–CTX nanoconjugate and HFn nanoparticles. The bar graph shows tM1 (CTX-AF488 signal vs HFn–AF647 signal, blue bars) and tM2 (HFn–AF647 signal vs CTX-AF488 signal, red bars) at the indicated time points.

Fig 3: Single-cell analysis of CD44 in liver cancer.UMAP plots of HCC (a) and iCCA (d) tumours derived from the single-cell RNA-seq datasets GSE166635 (HCC) and GSE138709 (iCCA), with major cell populations annotated (macrophages, T cells, and malignant cells). Feature plots showing CD44 expression levels across specific cell populations in HCC (b) and iCCA (e) samples (blue = high expression, grey = low expression). UMAP clustering of normal tissues (blue) versus tumour cells (orange) in an HCC sample (c) and an iCCA sample (f).

Fig 4: CD44 expression and patient survival in human HCC.a CD44 mRNA expression (TPM) in TCGA HCC tumours vs. adjacent normal liver (NTL). b CD44 mRNA levels by tumour stage (TCGA HCC). c Representative immunohistochemistry images of normal liver, adjacent non-tumorous liver, HCC, and iCCA tissues (images retrieved from the Human Protein Atlas). d Kaplan–Meier overall survival curves for TCGA HCC patients with CD44high vs. CD44low tumours. e Validation of survival difference in CD44high vs. CD44low groups using GEPIA2 (combined TCGA/GTEx data). ****=p < 0.0001.

Fig 5: Increase in CD44-positive hepatocytes with fibrosis progression.a Representative images of CD44 IHC in injury model livers (arrows indicate CD44-positive hepatocytes). b CD44v6 IHC in injury models (arrows indicate CD44v6-positive hepatocytes or cholangiocytes). c Quantification of CD44 and (d) CD44v6 expression based on integrated optical density (IOD) in the indicated liver injury models. e Representative images of liver sections from CCl4-treated mice at 4 weeks and 16 weeks, as well as an HCC. H&E staining, Sirius Red staining (fibrosis in red), and CD44 IHC (red arrows point to CD44-positive hepatocytes) are shown. f Quantification of CD44-positive hepatocytes in CCl4-treated mice over time. *=p < 0.05; **=p < 0.01.