Supplemental Figure S5.

Validation of immunofluorescence staining with FPR subtype-specific antibodies. Top: Comparison of immunofluorescence staining intensity of FPR-transfected HEK293T and U87 cells with receptor subtype specific antibodies. The hFPR1 (1 μg/ml), hFPR2 (0.2 μg/ml) and hFPR3 (1 μg/ml) antibodies were used. Note that the absolute staining intensities of all hFPRs in HEK293T cells were highly similar, however a strong reduction of hFPR2 staining in U87 cells is observed. No cross reactivity was detected. All images were acquired with the same settings and are displayed with the same illumination, brightness and contrast. Evidence for similar cell surface expression and production rates for hFPR1 and hFPR2 can be found in supporting Fig. S8. Bottom: corresponding nuclei staining with Hoechst 33342 demonstrating a comparable amount of total cell number in all visual fields.