The exosomal proteins were extracted by DNA/RNA/protein Isolation Kit (catalog number: R6734-02).
The sample was denatured by heating, separated by SDS-PAGE and transferred to a PVDF (polyvinylidene uoride) membrane. Next, the membranes were blocked with 5% skimmed milk powder and separately probed with rabbit anti CD9 (catalog number: 20597-1-AP, Proteintech), rabbit anti HSP70 (catalog number: GB11241, servicebio) and Mouse anti GAPDH (catalog number: 60004-1-Ig, Proteintech) overnight at 4°C with a nal dilution of 1:1000 (v/v). After three times of washing, the membranes were incubated with goat anti-rabbit secondary antibodies (catalog number: GB23303, Sevicebio) or goat antimouse secondary antibodies (catalog number: GB23301, Sevicebio) with 1:2000 dilution (v/v) at 37°C for 1.5 h. The images of membranes treated with ECL (enhance chemiluminescence) were captured by Western Blotting Detection System (Tiangen, Beijing, China).
The sample was denatured by heating, separated by SDS-PAGE and transferred to a PVDF (polyvinylidene uoride) membrane. Next, the membranes were blocked with 5% skimmed milk powder and separately probed with rabbit anti CD9 (catalog number: 20597-1-AP, Proteintech), rabbit anti HSP70 (catalog number: GB11241, servicebio) and Mouse anti GAPDH (catalog number: 60004-1-Ig, Proteintech) overnight at 4°C with a nal dilution of 1:1000 (v/v). After three times of washing, the membranes were incubated with goat anti-rabbit secondary antibodies (catalog number: GB23303, Sevicebio) or goat antimouse secondary antibodies (catalog number: GB23301, Sevicebio) with 1:2000 dilution (v/v) at 37°C for 1.5 h. The images of membranes treated with ECL (enhance chemiluminescence) were captured by Western Blotting Detection System (Tiangen, Beijing, China).