Fig. 5: TMEM233 is expressed in dorsal root ganglion neurons and can associate with NaV1.7.
From: Pain-causing stinging nettle toxins target TMEM233 to modulate NaV1.7 function
a Expression of tdTomato (red) and Nefh RNA, detected with RNAscope analysis (TS405, blue), in lumbar DRGs isolated from adult Tmem233Cre/Rosa-CAG-flox-stop-tdTomato mice. Inset, representative image of DRG section used for analysis, from 4 independent experiments. Scale bar, 200 μm. b Tmem233 and Scn9a RNA expression and localization in fresh-frozen sections of mouse DRGs determined by RNAscope analysis. Localization of Tmem233 mRNA (green, AF488) was compared to Nefh mRNA (red, Opal570) and Scn9a mRNA (far-red, Opal650, shown in magenta). Arrows, neurons expressing both Tmem233 and Scn9a leading to white signal color overlap. Scale bar, 100 μm. Image representative of 2 independent experiments. c SDS-PAGE gel of purified NaV1.7-TMEM233 complex. CBB (Coomassie brilliant blue staining), GFP (green fluorescent protein) fluorescence and mCherry fluorescence of the same SDS-PAGE gel. The green and red arrow indicate NaV1.7-GFP and mCherry-TMEM233 bands, respectively. MW, molecular weight marker (kDa). d Normalized size-exclusion chromatography profile of the purified NaV1.7-TMEM233 complex. Signals of total protein (black), GFP (green) and mCherry (red) were detected simultaneously. e Left: Quantification of proximity ligation assay signal in GFP-positive HEK293 cells transfected with GFP as well as N-terminal FLAG-tagged hNaV1.7 and N-terminal HA-tagged TMEM233 alone or in combination. Right: Representative images showing nuclei (DAPI, blue), GFP (Green) and proximity ligation signal (red channel, enlarged view in inset shown in grayscale for clarity). Scale bar, 10 μm. Data are shown as mean ± SEM; *, p < 0.05. n values and statistical information are detailed in Supplementary Table 1. Source data are included as a Source Data file.
