2.2.2 Co-localization of EP4 and COX-1
Dual-label immunocytochemistry was conducted according to the same procedure as mentioned previously in Section 2.4 and the DRG sections were incubated overnight at 4oC with the primary antibody for EP4 (1:200; Cayman Chemical ) and a COX-1 antibody (1:200; Cayman Chemical ) raised in mice. The DRG sections were then washed with PBS six times for 10 min each followed by incubation with the host-directed secondary antibody (EP4 primary, Texas Red-conjugated goat anti-rabbit, 1:500 dilution ; COX-1 antibody, Alexa-Fluor 488-conjugated goat anti-mouse, 1:200). The DRG sections were then washed again twice with PBS, mounted and photographed.