Rabbit anti cd133

For histochemistry or immunohistochemistry, the tissues were fixed in 4% paraformaldehyde (PFA), dehydrated in a series of graded ethanol solutions, embedded in paraffin, and cut into 5 μm sections using a rotary microtome (Leica, Germany). The sections were then used for subsequent H&E staining or immunofluorescence microscopy. The antibodies used for immunohistochemistry were rabbit anti-CD133 (1:100 dilution, Proteintech), rabbit anti-ALDH1A1(1:100 dilution, Proteintech), rabbit anti-ABCG2 (1:100 dilution, Cell Signaling Technology), rabbit anti-β-catenin (1:100 dilution, Proteintech), rabbit anti-SOD2 (1:100 dilution, Proteintech) and rabbit anti-GPX4 (1:100 dilution, Abcam). Images were analyzed using the ImageJ program.

Single cell suspensions were prepared and incubated with a rabbit anti-CD133 (1:50, Proteintech) for 30 min at 4 °C in dark. Cells were washed and then stained with FITC-conjugated goat anti-rabbit secondary antibody, followed by FACS analysis using a FACS Calibur™ flow cytometer (BD Immunocytometry Systems, Franklin Lakes, NJ). Dead cells were excluded by propidium iodide (PI) staining. The acquired data were analyzed with FlowJo vX.0.6 software (Tree Star Inc., Ashland, OR).