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Fig. 1. Expression profile of different cadherin subtypes in Xenopus cranial neural crest (CNC) cells. (A) cDNAs for real-time PCR quantification were synthesized using RNAs extracted from Xenopus CNC explants at stage 17, 20 and 23, respectively. (B) The identities of sample tissues were validated via RT-qPCR amplification with different tissue markers. CNC markers twist, slug and snail were present in high abundance in CNC samples throughout all three assayed stages. The mesodermal marker bra, the placodal marker eya1 and the epithelial marker k81 were barely detectable in all CNC samples. All values were normalized and calibrated to the expression of the reference gene ornithine decarboxylase (ODC) (expression of ODC=1). (C) Copy number (per cell) of E-, N-, XB/C-cadherin, Cadherin-11, PCNS and PAPC in CNC during migration. Copy number of each cadherin was calculated by absolute quantification using standard curves. At least three independent quantifications were performed and the bars indicate average values with standard deviations. Immunofluorescence staining of endogenous XB-cadherin (D), C/XB-cadherin (E), N-cadherin (F) and E-cadherin (G) on CNC explants. CNC cells were labelled with membrane GFP and explanted on a fibronectin-coated surface at stage 17. DAPI staining was used to visualize cell nuclei. All cadherins are prominently localized at cell–cell contacts. Scale bar: 10 μm. |
