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. 2023 Feb 17;33(4):273–287. doi: 10.1038/s41422-023-00782-7

Fig. 4. OLFR796 is a receptor of famsin.

Fig. 4

a Binding assay on frozen tissue sections from 8–10-week-old Olfr796+/+ and Olfr796–/– male mice. 100 nM GST or GST-famsin was incubated with frozen tissue slices. For competition binding, tissues were pretreated with 5 μM His-ProS2 or His-ProS2-famsin for 30 min. Scale bars, 20 μm. b Binding assay on mouse primary hepatocytes. 100 nM GST or GST-famsin was incubated with hepatocytes. For competition binding, cells were pretreated with 5 μM His-ProS2 or His-ProS2-famsin for 30 min. Scale bars, 10 μm. c Effect of WT OLFR796 or its mutant (Mut, R187D/R195D/E197A) on famsin binding in mouse primary hepatocytes. Data are shown as means ± SEM. Comparison of different groups was carried out using two-way ANOVA followed by Tukey’s test. ###P < 0.001. n = 3. d, e qPCR results (d) and in situ hybridization (e) showing relative mRNA levels of Olfr796 in different tissues from 8–10-week-old male mice. In the pancreas sections, the islets are outlined in red. OE, olfactory epithelium; OB, olfactory bulb. Data are shown as means ± SEM. n = 6 mice (d). f GST pull-down assay showing the interaction between GST-famsin and OLFR796-Flag or its mutant (Mut, R187D/R195D/E197A) in Hi-5 cells. g Quantification of the binding affinity between famsin and WT OLFR796 or its mutant (Mut, R187D/R195D/E197A) by MST. ΔFnorm indicates the change in the normalized fluorescence. Data are shown as means ± SEM. n = 3.