Extended Data Fig. 4: Expression of LRP8 variants in A549 cells and expression, purification and characterization of soluble LRP8 and PCDH1-EC1 proteins.
a, A549 cells expressing endogenous levels of LRP8 (WT) or cells overexpressing the indicated Flag-tagged LRP8 variants were immunostained with an anti-Flag mAb and visualized by fluorescence microscopy. Experiments were performed two times with similar results. Scale bar, 20 µm. b, A549 cells expressing endogenous levels of LRP8 (WT) or cells overexpressing the indicated Flag-tagged LRP8 variants were lysed and LRP8 expression levels were visualized by western blotting. The housekeeping protein β-actin was included as a loading control. Western blot images from one experiment representative of three independent experiments are shown. Mr, relative molecular weight (K denotes ×1,000). For gel source data, see Supplementary Information Fig. 3. c, Purified soluble LRP8-LA1-2, LRP8-LA1 and PCDH1-EC1 C-terminally fused to human IgG1 Fc tags were resolved on an SDS-polyacrylamide gel under non-reducing (- dithiothreitol, DTT) or reducing (+ DTT) conditions and visualized by Coomassie blue staining. Experiments were performed three times with similar results. Mr, relative molecular weight (K denotes ×1,000). For gel source data, see Supplementary Information Fig. 3. d, Analytical size-exclusion chromatography (SEC) of proteins from c. Absorbance (milli-absorbance units, mAu) was monitored at 280 nm.
