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. 2018 Feb 8;11:170–179. doi: 10.1016/j.omtn.2018.02.001

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© 2018 The Author(s)

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

PMC Copyright notice

CTS Promotes the Expression of miR-106a-5p through the Recruitment of PAX5 to the Promoter of miR-106a-5p

(A) Luciferase activity of different miR-106a-5p promoter constructs in cultured human chondrocytes with or without CTS stimulation. P1, promoter (−186 bp); P2, promoter (−439 bp); P3, promoter (−761 bp); P4, promoter (−1,000 bp); P5, mutated promoter (−761 bp). (B) Luciferase assay in cultured human chondrocytes infected with FLAG-tagged IRF2 or PAX5 or STAT4 and P5. Immunoblot shows the expression of IRF2, PAX5, and STAT4 with anti-FLAG. (C and D) Cultured human chondrocytes were infected with the PAX5 plasmid (C) and the vector control or shRNA for PAX5 (D), followed by stimulation with CTS as described in the Methods section. Immunoblot indicates the expression of PAX5. (E) Chromatin immunoprecipitation (ChIP) assay for PAX5 occupancy on the miR-106a-5p promoter or upstream of the promoter in cultured human chondrocytes. Each bar represents the mean ± SD of at least three independent experiments performed in triplicate. *p < 0.05, **p < 0.01.