Whole-mount fluorescent in situ hybridization was performed as previously described using the fgf10 probe(16 (link)). Whole mount IF staining was performed as previously described (16 (link)). Antibodies used include rabbit anti-Prox1a (1:100, GTX128354, GeneTex), goat anti-Hnf4a (1:50, sc6556, Santa Cruz), mouse anti-Alcama (1:20, zn-8, DSHB) and mouse anti-Islet1/2 (1:20, 39.4D5, DSHB), rabbit anti-pan-Cadherin (1:5000; Sigma), mouse anti-Anxa4 (1:100, ab71286, aka 2F11, Abcam)(15 (link)), rabbit anti-Bhmt (1:200, ab96415, Abcam), mouse anti-Mdr1(1:200, sc-71557, Santa Cruz), chicken anti-GFP (1:300, GFP1010, Aves Labs), goat anti-mCherry (1:500, LS-C204207, LSBio), rabbit anti-Cytokeratin7 (1:200, ab181598, Abcam), mouse anti-Sox9 (1:200, ab76997, Abcam). Imaging was performed on a Zeiss LSM710 running Zen 2010 (Black) and images were processed with Adobe Photoshop.
Protocol detail
Multimarker Fluorescent Imaging of Liver
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Antibodies
Aves
Bhmt
Cadherin 1
Chicken
Fgf10
Fluorescent in situ hybridization
Goat
Mouse
Rabbit
Sox9
Corresponding Organization : Sanford Burnham Prebys Medical Discovery Institute
Other organizations : Sichuan University, University of Cincinnati, Cincinnati Children's Hospital Medical Center, Morgridge Institute for Research, University of Wisconsin–Madison
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Variable analysis
independent variables
- Whole-mount fluorescent in situ hybridization using the fgf10 probe
- Whole mount IF staining
- Prox1a expression
- Hnf4a expression
- Alcama expression
- Islet1/2 expression
- Pan-Cadherin expression
- Anxa4 expression
- Bhmt expression
- Mdr1 expression
- GFP expression
- MCherry expression
- Cytokeratin7 expression
- Sox9 expression
- Experimental procedures described in previous publication (16)
- Not specified
- Not specified
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