JCS033928 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Non-SC laminins partially rescue the dysmyelinating phenotype in SC/DRG neuronal co-cultures from P0/Cre:fLAMγ1 mice after two weeks in myelinating promoting feed. (A) PCR analysis of genomic DNA of co-cultures from homozygous fLAM γ1 mice (F/F) and P0/Cre:fLAMγ1 mice (F/F, P0-Cre). The primers amplified the unrecombined (3.2 kb) and recombined (2.3 kb) fLAM γ1 alleles. (B) Myelination of SC/DRG co-cultures from fLAM γ1 mice eight days after addition of ascorbate (MF8) was detected by immunostaining for laminins (Ln; green) and MBP (red). (C and D) Myelination of SC/DRG co-cultures from P0/Cre:fLAMγ1 mice at MF8 (C) or MF14 (D) was detected by immunostaining for laminins (Ln; green) and MBP (red). Arrows indicate high level expression of laminins in the neuronal soma. Bar=50 µm.

• Supplemental Figure S2 -

  Fig. S2. Mouse SC/DRG neuronal co-cultures can be infected with Ad-eGFP-actin and Ad-mCherry-NFL and used for time-lapse imaging. (A) SC/DRG co-cultures were infected sequentially with Ad-mCherry-NFL at day 0 and Ad-eGFP-actin at day 10 in maintenance feeds. Images were taken and merged two days after the final infection. (B) Co-cultures were infected with Ad-mCherry-NFL and Ad-eGFP-actin, incubated for eight days in myelinating feed, and stained for MBP (blue). Infection of co-cultures with Ad-mCherry-NFL and Ad-eGFP-actin does not inhibit myelination. Note that the green SCs become elongated upon myelination. (C) Co-cultures infected with Ad-eGFP-actin were stained for S100 (red). Images were taken at green and red channel and merged. eGFP-positive cells colocalized with S100 immunostaining, indicating that these cells were SCs. Bar: (A,B) 50 µm; (C) 20 µm.

• Movie 1 -

  Movie 1. A control SC as it progressively changes shape to bipolar. Please refer to the legend of Fig. 2 for detailed information for producing Movie 1.

• Movie 2 -

  Movie 2. A laminin-deficient SC that exhibits no change in cell shape. Please refer to the legend of Fig. 2 for detailed information for producing Movie 2.

• Movie 3 -

  Movie 3. Process extension of a group of control SCs. Please refer to the legend of Fig. 3 for detailed information for producing Movie 3.

• Movie 4 -

  Movie 4. No process extension in laminin-deficient SCs. Please refer to the legend of Fig. 3 for detailed information for producing Movie 4.