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. 2022 Nov 11;11(22):3568. doi: 10.3390/cells11223568

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© 2022 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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Knockdown and re-/overexpression of YKL-40 in HCT116 and Caco2 cells. (A) Knockdown via CRISPR/Cas9 approach is described. Human YKL-40 is located on chromosome 1. The exon 3 sequence is reported, the gRNA complement sequence is underlined, and the PAM sequence is indicated with red nucleotides. (B) Gel image of Genomic Cleavage Detection Assay using DNA extracted from transfected HCT116 and Caco2 cells. Control template and primers were included as a PCR technical control. Cleavage efficiency is indicated for each sample. (C) Map of pCMV-CHI3L1-GFPSpark plasmid used for YKL-40 gene expression. Validation of YKL-40 knockdown and re-/overexpression at (D) mRNA and (E) protein levels in both cell lines. qPCR data and densitometric data are shown as mean ± SD. Statistically significant differences were calculated using Student’s t test: *, p < 0.001; **, p < 0.01; ***, p < 0.0001. Data represent three independent experiments.