JCS03016 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Phosphorylation of ERK upon NGF stimulation in adult DRG neurons. (A) When neurons were cultured on laminin, ERK phosphorylation (detected by phospho-ERK immunostaining) was significant enhanced upon NGF stimulation. (B) NGF treatment induced a similar increase in ERK phosphorylation of neurons cultured on poly-D-lysine. (C) Culturing neurons on aggrecan did not prevent ERK phosphorylation after NGF stimulation.

• Supplemental Figure 2 -

  Fig. S2. Axon assembly from adult naïve DRG neurons requires both neurotrophin and integrin signaling. (A) NGF (50ng/ml) induced extensive axon growth from adult naïve neurons plated on laminin. (B) Addition of the transcription inhibitor DRB (100 μM) at the time of plating neurons abolished NGF-induced axon growth. (C) Neurons primed with NGF for 7 hours grew axons in the absence of gene transcription. (D) Transcription-independent axon assembly was inhibited by NGF receptor TrkA inhibitor K252a (200nM). (E) NGF alone was unable to induce efficient axon assembly when neurons were plated on Poly-D-lysine (PDL) in the absence of laminin. (F) Quantification of axon assembly from adult naive neurons. % of neurons extending an axon over 24 hours is shown *P<0.0001 vs NGF+LM; ^#P<0.005 between indicated groups.

• Supplemental Figure 3 -

  Fig. S3. Cell adhesion molecule L1 did not support axon assembly of PCL neurons. When PCL neurons were cultured on L1-coated coverslips, little axon growth was observed.