Unboiled mouse tissue extracts (50 μg) were separated by 10% SDS-PAGE, and the membrane was blotted with VPAC2 antibody [HL3625] (GTX641600) diluted at 1:1000. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.

Boiled and unboiled rat tissue extracts (50 μg) were separated by 10% SDS-PAGE, and the membrane was blotted with VPAC2 antibody [HL3625] (GTX641600) diluted at 1:1000. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.

Unboiled RMS 13 whole cell and membrane extracts (30 μg) were separated by 10% SDS-PAGE, and the membrane was blotted with VPAC2 antibody [HL3625] (GTX641600) diluted at 1:1000. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody, and the signal was developed with Trident ECL plus-Enhanced.

Non-transfected (–) and transfected (+) boiled and unboiled 293T whole cell extracts (30 μg) were separated by 10% SDS-PAGE, and the membrane was blotted with VPAC2 antibody [HL3625] (GTX641600) diluted at 1:5000. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.