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Fig. 1 | BMC Genomics

Fig. 1

From: Transcriptional profiling reveals potential involvement of microvillous TRPM5-expressing cells in viral infection of the olfactory epithelium

Fig. 1

Fluorescence activated sorting (FACS) of cells isolated from the olfactory epithelium. a TRPM5 promoter driven expression of eGFP and OMP promoter driven expression of mCherry in the olfactory epithelium. Expression of eGFP is found both in MVCs that do not express mCherry (asterisk) and in OSNs double labeled with eGFP and mCherry (arrow). i. Composite, ii. eGFP, iii. mCherry, iv. Composite magnification. Magenta: mCherry, green: eGFP. Scale bar: i-iii, 50 μm, iv, 10 μm. b Schematic of RNA-seq process from tissue to RNA extraction. Mouse OE was dissociated into single cells and sorted via FACS. RNA was extracted from each of the resulting cell populations. c Two isolated OSNs differing in eGFP expression. Magenta: mCherry, green: eGFP. Scale bar: 10 μm. d Distribution of mCherry and eGFP fluorescence intensity for FACS-sorted cells. Three cell populations were isolated for RNAseq: Cells with low OMP promoter-driven mCherry expression and high TRPM5 promoter-driven eGFP expression (MVC_eGFP cells), cells with high OMP promoter-driven mCherry and low eGFP expression (OSN_eGFP- cells) and cells with eGFP expression of the same magnitude as MVC_eGFP cells and high OMP promoter-driven mCherry expression (OSN_eGFP+ cells). The number of cells collected for this FACS run were: OSN_eGFP-s 1,500,000, OSN_eGFP+s 5336 and MVC_eGFP cells 37,178. e qPCR levels (normalized to levels 18 s RNA) for expression of transcripts encoding for OMP (i), TRPM5 (ii), eGFP (iii) and ChAT (iv). The asterisks denote significant differences tested with either t-test or ranksum with p-values below the significance p-value corrected for multiple comparisons using the false discovery rate (pFDR) [15]. pFDR is 0.033 for OMP, 0.05 for TRPM5, 0.05 for eGFP and 0.03 for ChAT, n = 8 for OMP OSN_eGFP-s, 4 for OMP OSN_eGFP+s and 4 for MVC_eGFP cells

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