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Fig. 4. Suppression of axonal outgrowth by hnRNP K MO in intact animals. Embryos unilaterally injected with control MO (A), hnRNP K MO1 (B,D-H) or hnRNP K MO2 (C) processed at stage 37/38 by whole-mount immunofluorescence. (A-C) Embryos injected with control MO (A), hnRNP K MO1 (B) or MO2 (C), and immunostained for NF-M. Broken lines separate injected (lower) from uninjected (upper) sides. (B,C) Arrowheads on the injected side indicate peripheral motor axons; arrowheads on the uninjected side indicate residual wispy peripheral axons. (A) View under the fluorescence dissecting microscope; (B,C) stacked confocal optical sections. (D1) Neuronal β-tubulin staining in neuronal perikarya and peripheral axons of spinal cord, uninjected side. (D2) Tubulin staining in perikarya and a few scattered fibers, injected side. (E1-G2) Peripherin immunostaining of head (E1,E2) and spinal cord (F1,F2,G1,G2). (E1,F1,G1) Uninjected side; (E2,F2,G2) injected side. (E1,E2) Trigeminal ganglion neuronal perikarya (upper arrowhead) and nerves (lower arrowhead). (F1,F2) Spinal cord with peripheral motor axons. (G1,G2) Spinal cord neuronal perikarya. (H) Spinal cord immunostained for HNK-1. Robust staining of neuronal perikarya and axons on the uninjected side (below the broken line); clusters of stained neuronal perikarya (arrowheads) and very few fibers on the injected side (above the broken line). Scale bars in D1,E2,F1 and G2 also apply to D2,E1,F2 and G1, respectively. |
