(since image is not showing up in text body, see attached photo of my western blot results)
The first lane is the marker (ladder), and then the next lane is the control (which is not expressing any NTRK, so
there are no bands).
The left hand blot was probed with NTRK1 antibody and the blot on the right with NTRK2.
Lanes 2-5 show NTRK1 bands. All constructs are expressing properly and fairly equally.
Lanes 6-7 (on the right hand blot) are the WT NTRK2 and NTRK2 mutation, which are also expressing.
This confirms that all the genes are expressed.
The NTRK2 WT (lane 6) is expressed more than the NTRK2 mutation (lane 7) which actually provides more evidence to show that the NTRK2 mutation is transforming:
-Since the NTRK2 mutation grew out in 6% serum in the IL3 withdrawal assay and the WT did not, this blot suggests that it is not just
because the mutation was expressed at a higher level that this effect happened.
-Inferences: perhaps there is some loss of function in the WT after a certain amount of serum, while the NTRK2 mutation continues to thrive in that environment
I think I'd like to do the western blot again, but so far this looks good.