Anti mouse secondary antibodies

The protein extracts were separated by electrophoresis and transferred to polyvinylidene difluoride membranes (Millipore, Billerica, MA, USA). Membranes were blocked and incubated using the primary antibody anti-CD133 (Abcam, Cambridge, MA, USA), anti-CD44 (Abcam) and anti-β-actin antibody (Abcam). Then membranes were incubated with anti-mouse secondary antibodies (Abcam). Finally, protein bands were detected using Fluor Chem FC2 (Alpha Innotech, San Leandro, CA, USA) and their intensity was analyzed using the Image Lab software.

The protein extracts were separated by electrophoresis and transferred to polyvinylidene difluoride membranes (Millipore, MA, USA). Membranes were blocked and incubated using the primary antibody anti-CD133 (Abcam, MA, USA), anti-CD44 (Abcam) and anti-β-actin antibody (Abcam). Then membranes were incubated with anti-mouse secondary antibodies (Abcam) (7 (link)). Finally, protein bands were detected using Fluor Chem FC2 (Alpha Innotech, CA, USA) and their intensity was analyzed using the Image Lab software.

Protein concentrations were determined using a Bio-Rad DC Protein Assay kit (Bio-Rad Laboratories, Inc., Hercules, CA, United States). Equal amounts of protein were resolved on 10% SDS polyacrylamide gels and transferred to polyvinylidene fluoride (PVDF) membranes (EMD Millipore, Billerica, MA, United States). The membranes were subsequently incubated with rabbit anti-GRIA2 (1:100; Abcam, Cambridge, United Kingdom), rabbit anti-ENPP3 (1:1,000, ab190823, Abcam, Cambridge, United States), or mouse anti-β-actin (1:1000; A1978, Sigma Aldrich; Merck KGaA) overnight at 4°C. The next day, the membranes were incubated with the corresponding peroxidase-labeled goat anti-rabbit (1:10,000, Abcam, Cambridge, United States) or anti-mouse secondary antibodies (1:10,000, Abcam, Cambridge, United States). ImageJ software (National Institutes of Health, https://imagej.net/Citing) was used to quantify the densitometry of the bands.