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. 2024 Feb 20;15:1531. doi: 10.1038/s41467-024-45817-8

Fig. 7. SEL1L binds collagen as it is being synthesized via its FN2 domain and the FN2 domain is necessary for positive regulation of MRC2 expression by SEL1L.

Fig. 7

a Representative confocal immunofluorescence image of MRC5 fibroblasts, along with magnified inset to highlight co-localization, representative of N = 4 independent experiments. b Western blot of MRC5 fibroblasts after immunoprecipitation with anti-SEL1L antibody (vs. controls as indicated), representative of N = 2 independent experiments. c Western blot after immunoprecipitation with anti-procollagen type I antibody (with input controls as indicated) of HEK293T cells transfected with collagen and WT or mutant SEL1L constructs as indicated, representative of N = 3 independent experiments. d Representative confocal immunofluorescence images of HEK293T cells transfected with collagen and WT or mutant SEL1L constructs as indicated, representative of N = 3 independent experiments. e, f Western blot and densitometry of MEFs after treatment with WT or mutant SEL1L constructs as indicated, representative of N = 4 independent experiments; p = 0.0305 (EV vs. SEL1L), p > 0.9999 (EV vs. ΔFN2). g Flow-cytometry based measurement of MRC2 surface expression on MEFs after treatment with WT or mutant SEL1L constructs as indicated vs. Empty Vector (EV). N = 6 per group; p = 0.0244 (EV vs. SEL1L), p = 0.8913 (EV vs. ΔFN2). h–j Western blot and densitometry of HEK293T cells either WT or KO for SEL1L as indicated after treatment with EV (empty vector), WT or mutant SEL1L constructs as indicated, representative of N = 3 (i), 4 (j) independent experiments; p = 0.0011 (i, WT vs. EV), p = 0.0985 (i, WT vs. SEL1L), p = 0.5120 (i, WT vs. ΔFN2), p = 0.0241 (j, EV vs. SEL1L), p = 0.0028 (j, SEL1L vs. ΔFN2). k Q-RT-PCR of SEL1L-KO HEK293T cells after transfection with WT or mutant SEL1L as indicated vs. empty vehicle control. N = 3 per group; p = 0.0097 (KO vs. WT), p = 0.0036 (WT vs. ΔFN2), p = 0.6021 (KO vs. ΔFN2). l Comparison of protein domain structures of Sel1L orthologs across phylogenetically diverse organisms; FN2 Fibronectin-2 domain, tpr Tetratricopeptide repeat domain. Data are shown as the mean ± SEM. Statistics: f repeated-measures one-way ANOVA with post-hoc Bonferroni testing; g, k one-way ANOVA with post-hoc Tukey testing; i, j one-way ANOVA with post-hoc Bonferroni testing. *p < 0.05, **p < 0.01, ns not significant. Source data are provided as a Source Data file.