DEV022236 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Absence of hnRNP K from axons. (A-F) Transverse sections of optic nerve (A-C) and ventral spinal cord (D-F). Sections were stained by immunoperoxidase with diaminobenzidine/nickel chloride. Phase-contrast (A,D) and bright-field (B,E) views of sections stained for hnRNP K. Comparable sections are stained for MAP-1 (C) and a phosphorylated epitope of NF-M (F) to label axons. Ventral motoneurons are at the top of D-F. Scale bars: in A, 100 µm for A-C; in D, 50 µm for D-F, respectively.

• Supplemental Figure S2 -

  Fig. S2. Absence of any effect by Control MO or hnRNP E MO on hnRNP K expression. Two-cell embryos were unilaterally injected with a standard control MO (A) or an hnRNP E MO (B). Transverse optical sections obtained with the confocal microscope show stage 15 embryos processed for hnRNP K whole-mount immunostaining (red); green represents co-injected FITC-dextran. The vertical broken lines passing through the neural plate and notochord (top) separate the injected (left) and uninjected (right) sides.

• Supplemental Figure S3 -

  Fig. S3. Western blots illustrating suppression of hnRNP K and NF-M expressions by hnRNP K MO. Stage 29/30 embryos were homogenized and processed for chemiluminescent western blot using antibodies to hnRNP K, NF-M and GAPDH, as indicated. Lane 1, uninjected animals; lane 2, single-side injected with control MO; lane 3, unilaterally injected with hnRNP K MO1; lane 4, bilaterally injected with hnRNP K MO1. GAPDH staining of each respective blot was performed after staining for hnRNP K or NF-M. Graphs illustrate the intensities of each band normalized against the uninjected control (±s.d., three blots).