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. Author manuscript; available in PMC: 2010 Aug 1.
Published in final edited form as: Nat Genet. 2009 Dec 27;42(2):170–174. doi: 10.1038/ng.512

Figure 2.

Figure 2

Mutant TRPV4 causes neuronal toxicity. a) Quantification of TRPV4 transcript by qRT-PCR in control human lymphoblast lines (n=4), human dorsal spinal cord (n=3), ventral spinal cord (n=3), and tracheal cartilage (n=3) using qRT-PCR primers specific for following exon regions: exons 3–4, exons 5–6, exons 7–8, exon 5, and exon 7. Values are normalized to the first tracheal cartilage sample. Data is averaged; error bars, s.e.m. b) DRG neurons were transfected with WT and mutant forms of TRPV4 (green). At 16 hours, some cells expressing mutant forms of TRPV4 show evidence of early cellular toxicity with a collapsed cytoplasm. The nuclear DAPI stain is blue. Scale bar=40 µm. c) Quantification of propidium iodide uptake in DRG neurons expressing WT and mutant TRPV4 reveals a marked increase of cell toxicity in mutant expressing cells at 48 hours. *p<0.0001. d) HEK293 cells expressing R269C and R269H TRPV4 show an increase in the number of dead cells (red channel is EthD-1 stain) at 48 hours that is prevented by the TRP channel blocker ruthenium red (RR). Green channel is calcein-AM stain for live cells. e) Quantification of cell death in HEK293 cells indicates a time dependent increase in cell death that is blocked by the TRP channel blocker ruthenium red (RR). *p<0.01, **p<0.001. Data in d, f are averaged from three independent experiments; error bars, s.e.m.