Additional file 4 of Peptide targeting of lysophosphatidylinositol-sensing GPR55 for osteoclastogenesis tuning

Additional file 3. Figure S1. LPI-induced signalling is independent of GPR55 overexpression in HEK293T and HeLa cells. (a) FACS analysis with an anti-HA antibody of HEK293T cells transfected with empty vector (pcDNA3) or the vector coding for GPR55 wild-type (HA-GPR55) or mutants (as indicated). (b, c) Twenty-four hours after transfection, these HEK293T cells were serum deprived for 4 h, and then stimulated with 10 µM LPI for the indicated times. Western blotting for phosphorylated (p-ERK1/2) and total ERK1/2 are shown, from a representative experiment of three independent ones. (d). HeLa cells transfected with empty vector (pcDNA3.1) or the vector coding for the construct ss-3×HA-GPR55 (ssGPR55). Twenty-four hours after transfection, the cells were serum deprived for 2 h, and then stimulated with 10 µM soybean LPI for the indicated times. Western blotting for phosphorylated (p-AKT, p-ERK1/2) and total AKT and ERK1/2 are shown, from a representative experiment of three independent ones.