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. 1999 Sep 15;19(18):7889–7900. doi: 10.1523/JNEUROSCI.19-18-07889.1999

Fig. 2.

Fig. 2.

Neurite stimulation evokes a rapid nuclear response. A, Selective neurotrophin stimulation of DRG neurites results in pCREB immunostaining of DRG neuron cell bodies. After a 20 min neurotrophin (+NT) or vehicle control (−NT) (0.1 ng/ml BSA) stimulation at the neurites, DRG neuron cell bodies were fixed in 10% paraformaldehyde and immunostained using the anti-pCREB antibody in conjunction with a Cy3-conjugated secondary antibody. B, CREB phosphorylation is detected in the cell body within 5 min after selective neurotrophin (+NT) or vehicle control (−NT) (0.1 ng/ml BSA) stimulation at the neurites. Cell body lysates from nine compartmented cultures were pooled, separated on an SDS-PAGE gel, transferred to Immobilon membrane, and immunoblotted with anti-pCREB. C, Time course of CREB phosphorylation at the cell body in response to neurotrophin stimulation at the neurite. The intensity of the pCREB band was quantitated with the Molecular Dynamics Storm 860 imaging system using Image Quant (version 2.0; Molecular Dynamics), and normalized for protein loading. The time course of pCREB induction in response to neurotrophin stimulation at the neurites (calculated as the ratio between pCREB in neurotrophin and control-stimulated cultures) was calculated from several experiments (n = 4–10 for each time point; *p < 0.05). Diagram (top) is a schematic representation of neurons; black bar indicates presence of the Teflon divider. Area of DRG neurons analyzed is indicated by dashed lines.