Immunofluorescence Analysis of SMAD Signaling

Tissue sections were fixed in 4% paraformaldehyde and processed as previously described [18] (link). Immunofluorescence was performed in detail as previously described [19] (link). We used specific antibodies for pSMAD1/5/8 (Santa Cruz Biotechnology), total SMAD and ALK2 (both from Santa Cruz Biotechnology), ABCC6 (MRP6 antibody S-20, Santa Cruz Biotechnology), albumin (Bethyl Laboratories). The nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI) (Sigma–Aldrich) [19] (link).

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Blazquez-Medela A.M., Guihard P.J., Yao J., Jumabay M., Lusis A.J., Boström K.I, & Yao Y. (2015). ABCC6 deficiency is associated with activation of BMP signaling in liver and kidney. FEBS Open Bio, 5, 257-263.

Publication 2015

pSMAD1/5/8 total SMAD albumin 4′,6-diamidino-2-phenylindole (DAPI)

Abcc6 Albumin Alk2 Antibodies Antibody Immunofluorescence Nuclei Paraformaldehyde Tissue

Corresponding Organization : University of California, Los Angeles

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Variable analysis

independent variables

Tissue sections were fixed in 4% paraformaldehyde

dependent variables

Immunofluorescence for pSMAD1/5/8, total SMAD, ALK2, ABCC6 (MRP6), and albumin
Nuclei stained with DAPI

control variables

Tissue sections were processed as previously described [18]
Immunofluorescence was performed in detail as previously described [19]

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