Fig. 7: CHRNA5 binds to RABL6, primarily to the RABL6-39-279 region. | Cell Death Discovery

Fig. 7: CHRNA5 binds to RABL6, primarily to the RABL6-39-279 region.

From: Nicotine-induced activation of cholinergic receptor nicotinic alpha 5 subunit mediates the malignant behaviours of laryngeal squamous epithelial cells by interacting with RABL6

Fig. 7

The subcellular localisation of CHRNA5 and RABL6 proteins was examined in AMC-HN8 (A) and FD-LSC-1 (B) cells via fluorescence microscopy. Scale bar: 20 μm. C Representative images of molecular docking demonstrating the specific residues of the calculated binding site and combination types between CHRNA5 and RABL6. D Co-immunoprecipitation of endogenous CHRNA5 and RABL6. Whole-cell extracts from AMC-HN8 and FD-LSC-1 cells and the precipitates were analysed via SDS-PAGE, followed by western blotting with anti-CHRNA5/-RABL6 antibodies. IgG was used as the negative control. E GST pull-down assay indicated that recombinant GST-tagged CHRNA5, but not GST, interacted only with RABL6. F CHRNA5 specifically interacted with the RABL6-39-279aa region. Co-immunoprecipitation assay demonstrated that endogenous CHRNA5 and RABL6-39-279aa formed a complex in HEK293T cells. G Western blotting was performed to evaluate JAK2/STAT3 signalling-related proteins expression in LSCC cells infected with oe-NC and oe-RABL6 lentiviruses. H Proposed model of how nicotine exposure activates nicotinic acetylcholine receptors and consequently promotes the metastasis of LSCC.

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