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. 2022 Jun 21;8:58. doi: 10.1038/s41421-022-00402-5

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 5 — a Schematic representation of the experimental procedure. ASCs from patients (n = 3) were transfected with CLEC3B siRNA or scramble siRNA (negative control). The cells were harvested 48 h and 96 h after transfection for mRNA and protein analysis, respectively. b Images showing the ASCs after 96 h of transfection. Scale bar: 100 μm. c Network plot showing the downregulated pathways in ASCs with siRNA-mediated knockdown of CLEC3B versus the negative control. Functional enrichment analysis was performed on the differential expressed genes detected by bulk RNA-seq. Adjusted P value < 0.05, the hypergeometric test. d Significantly reduced mRNA expression of c3 markers and fibrosis-related genes in ASCs with CLEC3B knockdown versus the negative control (determined by bulk RNA-seq). The significance threshold was set to be a P value < 0.05 of the sleuth differential expression test. e Significantly reduced mRNA expression of c3 markers and fibrosis-related genes in ASCs with CLEC3B knockdown versus the negative control (determined by qPCR). The significance threshold was set to be a P value < 0.05 of the one-tailed paired t-test. f Immunoblot assays showing the decreased protein expression of CLEC3B, Collagen-1, SMAD2/3, and phosphorylated SMAD2/3 in ASCs with CLEC3B knockdown versus the negative control (n = 3). g Densitometry analysis of the immunoblot results. The significance threshold was set to be a P value < 0.05 of the one-tailed paired t-test. h Immunofluorescent staining of Collagen-1 in ASCs transfected with CLEC3B siRNA or scramble siRNA. Scale bar: 50 μm. Representative fields of view of ASCs from the patient L21 are shown. i Collagen-1 staining fluorescence intensity comparison. Each value represents the mean fluorescence intensity of three representative fields of view. Two-tailed paired t-test.