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. 2017 Jun 30;21(12):3494–3505. doi: 10.1111/jcmm.13260

Figure 2.

Figure 2

(A–C) Relative IL‐10 mRNA expression (A) and IL10 fourth intron luciferase activity (B/C) following LPS (10 ng/ml, 6 hr) was decreased through prior incubation with Akt/PKB kinase inhibitor Ly294002 (10 μM) in rMCs (A/B) and HUT78T cells (C). (D–F) Ex vivo ChIP assay revealed binding especially of p‐YB‐1S102 to the Il10 fourth intron in murine kidneys 12 hrs following LPS injection (1.5 mg/kg BW, i.p.). Used were anti‐YB‐1‐ (D/E) and anti‐p‐YB‐1S102‐(D/F) specific antibodies or unspecific IgG (negative control) with oligos within Il10 fourth intron region. Furthermore, amount of included DNA was tested without preceding immunoprecipitation (input). (G) Enhanced transcript numbers of IL‐10 in murine kidneys following LPS challenge. (H) mRNA expression of IL‐10 was increased in kidneys of both WT and Yb1 +/− animals 6 hrs after LPS injection (1.5 mg/kg BW, i.p.) compared with PBS control animals, but the increase was significantly dampened in Yb1 +/− mice (n = 5–6). (I/J) Enhanced YB‐1 expression resulted in elevated IL10 fourth intron luciferase activity in rMCs (I) and in T cells (HUT78) (J) following LPS challenge (10 ng/ml, 6 hr). Data are expressed as mean values ± S.D. *P < 0.05; **P < 0.01; ***P < 0.001.