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Figure 2.
A–G: Frontal views of the buccopharyngeal membrane labeled with pJNK1 antibody (green) at three different stages representing before and during perforation. White dots outline “holes” or perforations. H: Sagittal section through the buccopharyngeal membrane and surrounding face showing pJNK (green) and counterstained with propidium iodide. pJNK can also be observed in the two layered epidermis, in the outer epidermis (OE) and inner epidermis (IE). I: Flat mount of the epidermis labeled with pJNK antibody (green). J: The mesoderm labeled with pJNK antibody (green). J′) Corresponding image to J where the pJNK is labeled green and nuclei/DNA labeled with propidium iodide (red). White arrows indicate examples of cells that appear to be undergoing mitosis. K: Magnified image of a mesodermal cell labeled with pJNK antibody (green) (K′) corresponding cell from K labeled with propidium iodide to show DNA (red). K′: Cell corresponding to K showing a merge of pJNK (green) and DNA (red). L: Embryos labeled with pJNK antibody (red) and showing the fluorescein tagged control morpholino (green). L′: Same image as L showing only the pJNK labeling (red). M: Embryos labeled with pJNK antibody (red) and showing the fluorescein tagged JNK1 morpholino (green) M′: Same image as H showing only the pJNK labeling (red). pJNK1, phospho JNK1; st, stage. |
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Figure 6.
A: Schematic of the stages examined during buccopharyngeal perforation. B–G: Frontal views of the buccopharyngeal membrane labeled with E-cadherin in control and JNK inhibitor treated embryos. White arrows point to examples of cells with an increase in cytoplasmic labeling and white arrow heads indicate lower E-cadherin at the membrane. White dots outline the holes or perforation. H–M: Frontal views of the buccopharyngeal membrane labeled with clathrin in control and JNK inhibitor treated embryos. White dots outline the holes or perforation. N–S: Colocalization experiment with phospho-JNK (green) and clathrin (red). N: pJNK1 in green. O: Clathrin in red. P: Merge of pJNK1 and clathrin. Q: Volume rendered image of pJNK and clathrin co-labeling rotated 45 degrees. R: Scatterplot of pixels labeled in green vs. red in the image shown in P. S: Bar graphs of the average correlation coefficients for Pearson's and Mander's colocalization tests. T–X: Frontal views of the entire faces (regular letters) or only the buccopharyngeal membrane at two-fold magnification (prime letters) from representative embryos at stage 40–41. T,T′: Representative control embryo treated with DMSO. U,U′: Representative embryo treated with chlorpromazine. V,V′: Representative embryo treated with Bafilomycin. W,W′: Representative uninjected embryo. X,X′: Embryo injected with CA-JNK. Abbreviations: perf, perforated; cg, cement gland; CA-JNK, constitutively active JNK. Red dots outline the embryonic mouth. |
