Figure 2.

PXR and RXR interact and co-localise in human platelets. (A) RXR was immunoprecipitated from human washed platelets (8 × 10⁸ cells/ml) using a mouse monoclonal anti-RXR antibody. Immunoblot analysis was followed with the addition of a rabbit polyclonal anti-PXR antibody and its detection using a secondary antibody that does not recognize denatured IgG. The presence of RXR was also confirmed in the same samples. Cropped representative blot of 3 separate experiments is shown. Full length blot of PXR and RXR is shown in Supplementary Fig. 8Bi,ii respectively (B) Localisation of PXR and RXR in resting and activated (with 5 µM U46619 in the presence of integrelin) permeabilised human platelets was investigated using immunofluorescence microscopy. RXR (in red), PXR (in blue) and membrane GPIb receptors (in green) were stained using anti-RXR, anti-PXR and anti-GPIb antibodies respectively. Representative figures show the distribution of RXR and PXR in resting and activated platelets. (C) Scatter plots between the fluorescence intensity points of RXR and PXR in resting and activated platelets represent the degree of colocalisation between RXR and PXR. (D) The Pearson correlation coefficient (PCC) representing the degree of colocalisation between RXR-PXR in resting and activated platelets. PCC was quantified for 12 platelets using different fields. Data represent mean ± SD, **P < 0.01 and ***P < 0.001 was calculated by Student T-test. Figure adapted from corresponding PhD thesis⁴⁸.