Skip to main content
. 2021 May 26;297(1):100828. doi: 10.1016/j.jbc.2021.100828

Figure 4.

Figure 4

LL-34 enables U1 dsRNA association with scavenger receptor (SR) SRB1.A and B, NHEKs with gene knockdown for SRs SCARB1, SCARB1, SCARA3, CD36, LRP1, CD163L1, CXCL16, MSR1, MARCO, and OLR1 were cotreated with U1 dsRNA (2.5 μg/ml) and LL-34 (2 μM) for 18 h. Gene expression of IFNB1 and CXCL10 was assessed with quantitative RT-PCR (n = 3). C, proximity ligation assay (PLA) for U1 dsRNA and SR SRB1 binding. NHEKs were treated with LL-34-parent, LL-34-I24A, and LL-34-L31A (2 μM) for 15 min, followed by treatment with biotinylated U1 dsRNA (2 μg/ml) for 60 min at 4 °C. The physical proximity of LL-34 and SRB1 (C), and U1 dsRNA, and SRB1 (D) was determined with fluorescence-based PLA that produces a red fluorescent signal. DAPI (blue) was used as a nuclear counterstain. The scale bar represents 50 μm. Signal counts of C and D from five visual fields were used for PLA signal quantification, as shown in E and F (n = 3). (∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001). One-way ANOVA was used. DAPI, 4′,6-diamidino-2-phenylindole; LL-34, 34-amino acid peptide; NHEKs, normal human epidermal keratinocytes; ns, not significant.