Rabbit anti upar

LPS and EGF were purchased from Sigma-Aldrich, n-Octyl-D-glucoside was purchased from Santa Cruz Biotechnology. Primary antibodies were: mouse monoclonals Eno 276/3 and 72/1 against alpha-enolase^{19 (link)}; mouse monoclonal cmHsp70.1 against membrane Hsp70^{24 (link)}; goat anti-alpha-enolase, rabbit anti-E-cadherin, mouse anti-Hsp90, rabbit anti-Hsp70, goat anti-GAPDH, rabbit and mouse anti-EGFR, mouse anti-TFR, rabbit anti-uPAR, rabbit anti-Vimentin, goat anti-Lamin B (Santa Cruz Biotechnology); rabbit anti-phospho-Akt, mouse anti-Akt, rabbit anti-Phospho-p44/42 MAPK (Erk1/2) (Cell Signaling Technologies); mouse anti-ERK1/2 (Proteintech Group), mouse anti-β-actin (Sigma-Aldrich); rabbit anti-TLR4 (Novus Biologicals); rabbit anti-Caveolin-1 (Upstate Biotechnology); mouse anti-Annexin-A2 (BD Biosciences). Alexa Fluor-conjugated secondary antibodies were purchased from ThermoFisher Scientific Inc.

Lysates of MVs and their parent MSCs harvested under hypoxia or normoxia (50 µg) were electrophoresed through a 12% SDS-polyacrylamide gel. The proteins were then transferred to a PVDF membrane at 200 mA for 2 h, after which Western blot analysis was conducted, After blocking with 5% skim milk, the membrane was subsequently incubated with 1∶400 diluted rabbit anti-UPAR, 1∶200 diluted mouse anti-Angiogenin, or 1∶1000 diluted mouse anti-GAPDH (Santa Cruz Biotechnology, Santa Cruz, USA) and washed three times for 10 min with TBST buffer. Next, the membrane was incubated with HRP conjugated anti-rabbit or anti-mouse IgG. After being washed three times with TBST buffer for 10 min, the HRP activity was observed by applying a chemiluminescent substrate (Applygen Technologies In) and exposing the samples to an X-ray film.