Extended Data Fig. 10: Validation of the anti-human Gremlin1 antibody for various applications and gating strategy for flow cytometry in this study.
(a-c) Antibody specificity of the anti-human Gremlin1 antibody (Mabspace Biosciences) in immunoprecipitation, immunoblotting, IHC and IF was validated using GREM1 overexpressing, GREM1 knockout and respective control LNCaP cells. IHC Scale bars = 50 μm, IF Scale bars = 20 μm. The immunostaining experiment was repeated at least twice and representative images were presented. The immunoblotting experiment was repeated at least 3 times with similar results. (d) Mass spectrum of immunoprecipitants with the anti-human Gremlin1 antibody (Mabspace Biosciences) in LNCaP cells shows an enrichment of the Gremlin1 protein. The experiment was repeated at least 3 times with similar results. (e) Gating strategy for the flow cytometric examination of apoptosis in this study. The frequency of the cells in the apoptotic stage were calculated based on APC + /DAPI−.
