RAGE interacts with FPR1 and FPRL1 in glial and transfected HEK293 cells. (A) Membrane proteins from astrocytes or microglia were extracted using anti-ratFPRL1 or ratFPR1 antibodies. (B) Membrane proteins from FPR1 or FPRL1 and RAGE or ΔRAGE transfected HEK293 cells were extracted and anti-humanFPRL1 or humanFPR1 antibodies were used to precipitate. The resulting immunoprecipitates were electrophoretically separated, transferred to nitrocellulose and detected with anti-FPR1, anti-FPRL1 and anti-RAGE antibodies. FPR1 and FPRL1 are co-immunoprecipitate with RAGE in glial as well as transfected HEK293 cells. The positions of molecular mass markers are indicated on the left (in kDa). The values representing mean ± SD of protein and phosphorylation levels derive from densitometric quantification of three independent experiments for co-immunoprecipitated RAGE (C for glial cells and D for transfected HEK293 cells) normalised to FPR1 or FPRL1. An asterisk indicates a significant difference (*p < 0.05) compared to controls on the basis of one-way ANOVA and Bonferroni post-hoc tests. (E) Western Blotting for different receptors of whole cell lysate from astrocytes, microglial and transfected HEK293 cells (each 10 μg protein). The positions of molecular mass markers are indicated on the left (in kDa) and the target on the right.