Fig. S4
The GFP-positive cells are NaR cells. a) GFP-positive cells express trpv5/6 mRNA. Tg(igfbp5a:GFP) raised in normal [Ca2+] medium were sampled at 120 hpf and analyzed by in situ hybridization for trpv5/6 mRNA expression (green, left panels) and GFP immunostaining (red, middle panel). Merged view is shown in the right panel. Scale bar = 50 μm. b-c) The igfbp5a mRNA (b) and trpv5/6 mRNA (c) is highly expressed in GFP-positive cells . Tg(igfbp5a:GFP) larvae (72 hpf) were transferred to 0.2 or 0.001 mM [Ca2+] medium. 18h later, GFP-positive cells were isolated by FACS as described in Materials and Methods. The levels of igfbp5a and trpv5/6 mRNA were determined by qPCR and normalized by the 18s mRNA levels. Values are mean ± SEM of two independent experiments. GFP+ and GFP- indicates GFP positive- and GFP-negative cells. |
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Stage: | Day 5 |