Supplementary material

Video 1: Expression of CD44 in tumor cells. Tissue sections of clear cell renal cell carcinoma were obtained from Sun Yat-sen University Cancer Center. The slides were deparaffinized and rehydrated with distilled water. Following rehydration, the slides were soaked in citrate buffer, boiled for 15 min, gradually cooled down to room temperature, and then washed with PBS for 5 min. After three washes, the tissue sections were circled with PAP-PEN and blocked with 3% BSA for 30 min at room temperature. Subsequently, they were stained with rabbit anti-CD44 antibody (GB113500, SERVICEBIO) at 1:200 dilution at 4 °C overnight. The next day, the slides were washed three times with PBS and incubated with Alexa Fluor 488-conjugated anti-rabbit IgG (GB25303, SERVICEBIO) at 1:400 dilution at room temperature for 50 min. After three washes in PBS, cell nuclei were stained with DAPI (G1012, SERVICEBIO) for 10 min at room temperature. All slides were mounted with antifade mountant (G1401, SERVICEBIO). The fluorescence images were obtained using a NIKON Eclipse Ti confocal microscope (Nikon Instruments) and reconstructed in 3D using NIS-Elements Viewer 5.2 software (Lim-Instruments/Nikon Instruments) with a volume of 637.08 × 637.08 × 19 μm³. The sequence of selected 50 images was then converted into a video.

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© The Authors, published by EDP Sciences, 2024