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. 2023 Jan 3;24(1):875. doi: 10.3390/ijms24010875

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© 2023 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Stimulation by LL-37 alone or in combination with poly(I:C) in NHEKs is mediated by scavenger receptors. SCARA3 (SR-A3), SCARB1 (SR-B1), CD68 (SR-D1), OLR1 (SR-E1), AGER (SR-J1), and non-targeting pool siRNAs were transfected into NHEKs, and the cells were stimulated with LL-37, poly(I:C) or LL-37+poly(I:C) 2 days later. After 6 h of incubation, mRNA induction was evaluated by RT-PCR. The inductions of (a) PTGS2, (b) VEGFA, (c) CXCL10, (d) IFNB1, and (e) TNFA are shown, respectively. (f) PLA was performed for various scavenger receptors and LL-37. LL-37 was added to NHEKs, 10 min later poly(I:C) was added, and the cells were incubated at 4 °C for 1 h. Using antibodies against various scavenger receptors and LL-37, the proximity of each scavenger receptor and LL-37 was detected by fluorescence-based PLA as green fluorescent spots. Nuclei (blue) were counterstained with DAPI. Data are means ± SEM of three biological replicates. ** p ≤ 0.01, **** p ≤ 0.0001 by two-way ANOVA with Bonferroni’s post hoc test. See also Figures S3 and S4.

Stimulation by LL-37 alone or in combination with poly(I:C) in NHEKs is mediated by scavenger receptors. SCARA3 (SR-A3), SCARB1 (SR-B1), CD68 (SR-D1), OLR1 (SR-E1), AGER (SR-J1), and non-targeting pool siRNAs were transfected into NHEKs, and the cells were stimulated with LL-37, poly(I:C) or LL-37+poly(I:C) 2 days later. After 6 h of incubation, mRNA induction was evaluated by RT-PCR. The inductions of (a) PTGS2, (b) VEGFA, (c) CXCL10, (d) IFNB1, and (e) TNFA are shown, respectively. (f) PLA was performed for various scavenger receptors and LL-37. LL-37 was added to NHEKs, 10 min later poly(I:C) was added, and the cells were incubated at 4 °C for 1 h. Using antibodies against various scavenger receptors and LL-37, the proximity of each scavenger receptor and LL-37 was detected by fluorescence-based PLA as green fluorescent spots. Nuclei (blue) were counterstained with DAPI. Data are means ± SEM of three biological replicates. ** p ≤ 0.01, **** p ≤ 0.0001 by two-way ANOVA with Bonferroni’s post hoc test. See also Figures S3 and S4.