ZFIN Image: Xue et al., 2014, Fig. S5

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Fig. S5

Bmp2b and p-Smad1/5/8 levels appear unaffected in earlier embryos depleted of organizer Bmp signals. One-cell embryos were injected with 105 pg Tg(-2.067gsc:tBr-gfp) or Tg(-2.272bmp2b:tBr-gfp) plasmid DNA and fixed at 40% and 55% epiboly stages. Uninjected or injected embryos were immunostained using anti-Bmp2b or anti-p-Smad1/5/8 antibody along with DAPI staining. The immunostained embryos were imaged by confocal microscopy from the animal pole. A series of optical equatorial sections at an interval of 3.5 μm were obtained, but only 2 or 3 sections at the blastodermal margin were used for intensity estimation. Eight areas on half of each section (as illustrated above curves), 323.28 μm² each, were chosen to evaluate immunostaining intensity and DAPI intensity. For each group, 3-5 embryos were analyzed. The ratio of immunostaining intensity to DAPI represented relative immunostaining intensity, and the ratio in the ventralmost position was set to 1.0. (a-d) Representative confocal images and average relative immunostaining intensities. The standard deviations were indicated.

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