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. 2021 Jan 25;64:103213. doi: 10.1016/j.ebiom.2021.103213

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© 2021 The Authors

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

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Fig 5 — IL-24 exerted protective effects on HSCs and hepatocytes in an IL-20R1-independent manner. The effects of IL-24 or IL-20 on the isolated primary HSCs from wild-type, IL-20R1-, and IL-20R2- deficient mice were analyzed. (a) Primary HSCs were isolated for cell culture. PBS or IL-24 were added to the cells on day 5. Two days later, mRNA was harvested and was analyzed for the gene expression of the activated HSCs marker – α-SMA (Acta2) by real-time PCR with specific primers. Gapdh was used as an internal control. Two-tailed unpaired t-test, p = 0•0002*** between WT+PBS and WT+IL-24, p = 0•0012** between IL-20R1KO+PBS and IL-20R1KO+IL-24, p = 0•6744 between IL-20R2KO+PBS and IL-20R2KO+IL-24. (b) The effects of IL-20 on the gene expression of Acta2 was also analyzed. Two-tailed unpaired t-test, p = 0•0002*** between WT+PBS and WT+IL-20, p = 0•0926 between IL-20R1KO+PBS and IL-20R1KO+IL-20, p = 0•7326 between IL-20R2KO+PBS and IL-20R2KO+IL-20. (c) The effect of IL-24 on the gene expression of Pcna in primary HSCs was analyzed by real-time PCR. Two-tailed unpaired t-test, p = 0•0329* between WT+PBS and WT+IL-24, p = 0•0208* between IL-20R1KO+PBS and IL-20R1KO+IL-24, p = 0•5565 between IL-20R2KO+PBS and IL-20R2KO+IL-24. (d) The effects of IL-20 on the gene expression of Pcna in primary HSCs. Two-tailed unpaired t-test, p = 0•0117* between WT+PBS and WT+IL-20, p = 0•4168 between IL-20R1KO+PBS and IL-20R1KO+IL-20, p = 0•6304 between IL-20R2KO+PBS and IL-20R2KO+IL-20. (e) Primary hepatocytes isolated from WT, IL-20R1-, and IL-20R2-deficient mice were treated with PBS or IL-24 for 8 h. The mRNA transcript of Tgfb was analyzed by real-time PCR. Two-tailed unpaired t-test, p = 0•0373* between WT+PBS and WT+IL-24, p = 0•0465* between IL-20R1KO+PBS and IL-20R1KO+IL-24, p = 0•6684 between IL-20R2KO+PBS and IL-20R2KO+IL-24. (f) The effects of IL-20 on the gene expression of Tgfb were analyzed. Two-tailed unpaired t-test, p = 0•0012** between WT+PBS and WT+IL-20, p = 0•1702 between IL-20R1KO+PBS and IL-20R1KO+IL-20, p = 0•1911 between IL-20R2KO+PBS and IL-20R2KO+IL-20. (g) PBS or IL-24 was added to shScramble, shIL-20R2, and shIL-22R1 AML12 mouse hepatocytes cell line for 6 h. The mRNA transcript of Col1a1 was analyzed by real-time PCR. Two-tailed unpaired t-test, p = 0•0089** between WT+PBS and WT+IL-24, p = 0•05,799 between IL-20R1KO+PBS and IL-20R1KO+IL-24, p = 0•9279 between IL-20R2KO+PBS and IL-20R2KO+IL-24. (h) The effects of IL-20 on the expression of Col1a1 in shScramble, shIL-20R2, and shIL-22R1 AML12 cells were analyzed. Two-tailed unpaired t-test, p = 0•0012** between WT+PBS and WT+IL-20, p = 0•1088 between IL-20R1KO+PBS and IL-20R1KO+IL-20, p<0•0001**** between IL-20R2KO+PBS and IL-20R2KO+IL-20. Data are means ± SEM. The experiments in a-h were repeated three times independently with similar results, and the data of one representative experiment was shown.