Skip to main content
. 2010 Feb 15;120(3):850–858. doi: 10.1172/JCI41013

Figure 3. NT-3/TrkC interference promotes TrkC proapoptotic activity.

Figure 3

(A) CLB-Ge2 cells were transfected with either empty vector or with a plasmid encoding the dominant-negative TrkC-IC D641N and treated for 24 hours with or without anti-TrkC blocking antibody Cell death was monitored by TUNEL labeling of cells plated on slides. The control panel shows TrkC-IC D641N by Western blots using anti-TrkC antibody (bottom panel). Representative images are shown. Original magnification, ×20. (B) The efficacy of TrkC siRNA was evaluated by Western blot on nonexpressing TrkC 13.S.24 olfactive neuroblasts. Cells were transfected either with empty vector or with uncleavable TrkC D945N D641N double mutant that does not trigger apoptosis, and with scrambled siRNA or TrkC siRNA (siRNA TrkC). (C) Cell death induction in the CLB-Ge2 cell line was quantified after transfection with either scrambled siRNA (siScr), TrkC siRNA (siTrkC), NT-3 siRNA (siNT-3), or a mix of TrkC and NT-3 siRNA, using relative caspase-3 activity assay. (D) Phospho-Akt and phospho-Erk levels of CLB-Ge2 cells were monitored by Western blot after 16 hours of treatment with 2 μg/ml anti-TrkC blocking antibody 20 nM Ly29402, 100 nM U0126, or 100 ng/ml NT-3, in absence of serum. (E) Detection of TrkC cleavage band (20 kDa, indicated by the arrow) by Western blot, using an anti-TrkC antibody on cells treated (or not) with anti-TrkC blocking antibody, with or without the general caspase inhibitor BAF. (A and C) Data represent mean ± SEM. *P < 0.05, 2-sided Mann-Whitney test, compared with control.