Figure 2. SELENOP is expressed in colonic macrophages and modifies migration.

(A) Selenop mRNA was detected by RNAscope. Negative control probe shown on left. Inset areas marked by dotted lines. Scale bar = 50μm. (B) qPCR of Selenop expression in BMDM (n=3) and whole colon tissue (n=3) as compared to liver (n=4) and brain tissue (n=4). Selenop levels were normalized to Gapdh and represented as fold change over liver expression. (C) Representative image (left) and quantification (right) of multiplex RNAscope for Selenop (green) and Cd68 (red) in murine colon tissue. Results represented as percent co-labeled Selenop (+) stromal cells per high powered field (n=15). Scale bar = 25μm. (D) Selenop expression was assessed by qPCR in BMDM established from Selenop^WT, Selenop^ΔMye/+, and Selenop^ΔMye mice (n=2/genotype with 3 technical replicates). (E) BMDM were plated on transwell filters and allowed to migrate over 72 hours. Representative images (left) and quantification (right). For quantification, the number of migrated cells was assessed over 5 low power images per well and combined to yield the number of migrated cells. Results are combined data points from 3 independent experiments, each containing 3 technical replicates, represented as fold change over Selenop^WT. (F) qPCR analysis of matrix-associated genes, biglycan/Bgn, transgelin/Tagln, and tissue inhibitor of metalloproteinases 3/Timp3 and (G) Gpx1 and Gpx3 (n=4/genotype with 3 technical replicates). (H) GPx1 protein assessed from BMDM. Representative results (left) and quantification normalized to tubulin and represented as fold change over Selenop^WT (right) (n=5 Selenop^WT, 4 Selenop^ΔMye/+, and 5 Selenop^ΔMye). *P<0.05, **P<0.01, ****P<0.0001, Student’s t test (C) or one-way ANOVA with Tukey’s multiple comparisons test (D-H). For all qPCR analysis, results were normalized to Gapdh and represented as fold change over Selenop^WT.