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

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© The Author(s) 2024

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — a Heatmap of overall phenotype scores from original screens as indicated for top genes from ‘Collagen Biosynthetic Process’ GO gene-set (displayed genes had most extreme ‘Score’ values in one or more screen result, with most extreme ‘Score’ absolute value at least >1 and concordant with GO term; 29/43 genes from this GO term that were included in the screen fit these criteria and are displayed). b Flow-cytometry based collagen-uptake assay in U937 cells after shRNA treatment as indicated vs. Scramble control. MFI Mean Fluorescence Intensity. N = 4 per group; p < 0.0001. c Western blot of U937 cells after shRNA treatment as indicated vs. Scramble control, representative of N = 4 independent experiments. d Q-RT-PCR in U937 cells after shRNA treatment as indicated vs. Scramble control. N = 4 per group; p = 0.0483. e, f Western blot and densitometry of MRC5 fibroblasts after shRNA treatment as indicated vs. Scramble control, representative of N = 3 independent experiments; p = 0.0477. g Representative confocal immunofluorescence images of MRC5 fibroblasts after shRNA treatment as indicated vs. Scramble control. h Flow-cytometry based collagen-uptake assay in MRC5 cells after shRNA treatment as indicated vs. Scramble control. N = 4 per group; p < 0.0001. i Flow-cytometry based collagen-uptake assay in U937 cells after shRNA treatment as indicated vs. Scramble control. N = 6 (Scramble), 4 (shMRC2, shCOL1A1, shCOL5A1), 5 (shCOL1A2, shCOL5A3) independent biological replicates; p-values are for post-hoc testing of Scramble vs. the following: shMRC2 (p < 0.0001), shCOL1A1 (p = 0.0005), shCOL1A2 (p = 0.0088), shCOL5A1 (p = 0.0006), shCOL5A3 (p = 0.0001). (j–k) Q-RT-PCR in U937 cells after shRNA treatment as indicated vs. Scramble control. N = 5 (left, all groups), 4 (right, Scramble, shCOL5A1, shCOL5A3), 5 (right, shMRC2) independent biological replicates; j p = 0.0136 (Scramble vs. shMRC2), 0.0189 (Scramble vs. shCOL1A1), 0.0041 (Scramble vs. shCOL1A2); k p = 0.0080 (Scramble vs. shMRC2), 0.0223 (Scramble vs. shCOL5A1). l Flow-cytometry based collagen-uptake assay in MRC5 fibroblasts after shRNA treatment as indicated vs. Scramble control. N = 6 and 4 in Scramble and shCOL1A1 groups respectively, N = 2 in shMRC2 group; p = 0.0011. m, n Western blot and densitometry of MRC5 fibroblasts after shRNA treatment as indicated vs. Scramble control, representative of N = 4 independent experiments; p = 0.0137 (left), p = 0.0017 (right). Data are shown as the mean ± SEM. Statistics: b–k one-way ANOVA with post-hoc Dunnett’s testing; (l–n) unpaired Student’s t test (two-sided). *p < 0.05, **p < 0.01, ****p < 0.0001. Source data are provided as a Source Data file.