Figure 3
Expression of NaV1.6 channels in human cervical cancer cell lines. (A) Immunofluorescence confocal microscopy analysis of NaV1.6 channels expression in C33A, SiHa, HeLa, and HEK293 cells stably expressing NaV1.6 channels (HEK-Nav1.6; positive control). Cervical cancer cells were incubated with an antibody against NaV1.6 protein followed by a staining with FITC-coupled secondary antibody. DAPI reagent was used for nucleus staining. Image acquisition was performed each 0.33 µm in a total thickness of 6.5 µm. Confocal sections were merged and 3D-reconstructions were performed from Z-planes for each region of interest. Orthogonal projections from xz and yz planes of confocal images show positive signal for NaV1.6 protein (far right panel), indicated by red arrows in xz and yellow arrows in yz planes, respectively. Scale bar, 10 µm. (B) Western blot analysis of NaV1.6 channel protein in total (T), cytoplasmic (C) and nuclear (N) protein extracts from human cervical cancer cell lines. Histone-3 and GAPDH proteins were used to demonstrate the enriching of subcellular fractions. Representative blot of three independent experiments. Notice that a ~150 kDa anti-NaV1.6 reactive protein (red arrows) was found in nuclear fraction from cancer cells. The full-length ~250 kDa NaV1.6 protein was only observed in total and cytoplasmic protein extract from C33A cells.
