Characterization Of Bitter Taste Receptor Dependent Autophagy in Oral Epithelial Cells

Abstract

Microbial dysbiosis is an important trigger in the development of oral diseases. Oral keratinocytes or gingival epithelial cells (GECs) offer protection against various microbial insults. Recent studies suggest GECs expressed higher level of bitter taste receptor 14 (T2R14) compared to other taste receptors and toll-like receptors and acts as innate immune sentinels. Macroautophagy or autophagy is a cellular conserved process involved in the regulation of host innate immune responses against microbial infection. Here, we describe a robust method for evaluation of T2R14-dependent autophagy flux in GECs. Autophagy flux was detected using western blot analysis in GECs and further was confirmed using Acridine Orange dependent flow cytometry analysis. Workflow O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=151 SRC="FIGDIR/small/578576v1_ufig1.gif" ALT="Figure 1"> View larger version (21K): org.highwire.dtl.DTLVardef@1fa7b46org.highwire.dtl.DTLVardef@1b333baorg.highwire.dtl.DTLVardef@939d5org.highwire.dtl.DTLVardef@1b3f6f9_HPS_FORMAT_FIGEXP M_FIG C_FIG Graphical abstractSchematic showing the methodology (Western blot and flow cytometry) used for assessment of autophagy flux in GEC (created with Biorender). GEC: Gingival epithelial cells, BafA1: Bafilomycin, Rapa: Rapamycin, LC3-II: microtubule associated light chain protein, p62: sequestosome 1