To determine the effect of CDK5 on wildtype and S17A mutant spinophilin, the fluorescence intensity for NF-M in the HA (spinophilin) immunoprecipitate was normalized to fluorescence intensity of spinophilin in the HA IP.
This value was based on the decrease in NF-M protein isolated from spinophilin immunoprecipitates in lesion compared to intact striatum (see below).
NF-M Is Decreased in Spinophilin IPs after 6-OHDA Lesion.
NF-M Coimmunoprecipitates with Spinophilin and PP1.
The mechanisms by which the association between spinophilin and synaptic proteins such as NF-M may be modulated following DA depletion are not known.
The Interaction of Spinophilin and NF-M in HEK293 Cells Requires Overexpression of the PKA Catalytic Subunit.
When reacted with SMI 31 antibody; protein bands corresponding to NF-H and NF-M were not seen at 2 months interval.
The SMI 31 antibody reactive NF-H and NF-M bands were observed at 2 and 4 months, but were absent at 6 and 12 months.
The presence of NF-H and NF-M proteins was revealed at 4 months interval (Figures 5a and 5b); but at the subsequent intervals of 6, 12, 16 and 20 months SMI 31 reactive NF-H and NF-M bands were not seen (Figure 5a).
It is interesting to note that NF-H, NF-M and NF-L bands were seen at 4 months.
smegmatis demonstrated protein bands corresponding to NF-H and NF-M from 4-20 months post inoculation.
Decreased immunoreactivity to SMI 31 antibody and faint or absent NF-H and NF-M protein bands in both viable and heat killed M.