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. 2021 Jan 1;11(1):426–444. doi: 10.7150/thno.50281

Figure 1.

Figure 1

Platr4 is an oscillating and steatohepatitis-related lncRNA whose expression is driven by NF-κB. (A) Representative micrographs of Oil Red O, H&E and Sirius red staining of liver sections from normal mice and mice with MCD-induced steatohepatitis. Scale bar = 50 μm. (B) Heatmap diagram for differentially expressed (DE) genes in livers from normal mice and mice with MCD-induced steatohepatitis. (C) Venn diagram showing the numbers of differentially expressed lncRNAs (DE lncRNAs) in the livers of wild-type and Rev-erbα-/- mice at ZT6 and ZT10. Data are available upon request. (D) Diurnal hepatic Platr4 expression in normal mice and mice with MCD-induced steatohepatitis. Data are mean ± SD (n = 8). *p <0.05 versus normal mice at individual time points (two-way ANOVA and Bonferroni post hoc test). (E) Expression comparisons of Platr4 in the various tissues and BMDMs of wild-type mice. Data are mean ± SD (n = 3). (F) Expression comparisons of Platr4 in BMDMs, hepatic stellate cells (HSCs), hepatocytes and Kupffer cells (KCs) derived from wild-type mice. Data are mean ± SD (n = 3). (G) Relative cytoplasm and nuclear expression of Platr4 in Kupffer cells and BMDMs of wild-type mice. Data are mean ± SD (n = 3). (H) FISH analysis of subcellular Platr4 (green) location in BMDMs from wild-type mice. (I) Effects of Pam3CSK4 (a TLR1/2 agonist, 100 nM), Poly (I:C) (a TLR3 agonist, 25 µg/ml) and LPS (a TLR4 agonist, 500 ng/ml) on Platr4 expression in BMDMs. Data are mean ± SD (n = 3). *p <0.05 versus CTRL (one-way ANOVA and Bonferroni post hoc test). (J) Dose-dependent effects of LPS (0.5 and 1 μg/ml) on Platr4 and IL-1β mRNA in BMDMs. Data are mean ± SD (n = 3). *p <0.05 (one-way ANOVA and Bonferroni post hoc test). (K) Effects of LPS (500 ng/ml) on Platr4 expression in synchronized BMDMs. Data are mean ± SD (n = 3). *p <0.05 versus control at individual time points (two-way ANOVA and Bonferroni post hoc test). (L) Effects of Bay 11-7082 on Platr4 and IL-1β mRNA in unstimulated or LPS-stimulated BMDMs. Data are mean ± SD (n = 3). *p <0.05 (two-way ANOVA and Bonferroni post hoc test). (M) Dose-dependent effects of p65 plasmid (0.75 or 1 μg) on Platr4 expression in BMDMs. Data are mean ± SD (n = 3). *p <0.05 (one-way ANOVA and Bonferroni post hoc test). (N) Dose-dependent effects of p65 plasmid (0.25, 0.5 or 1 μg) on 2.1 kb Platr4 reporter activity in BMDMs. Data are mean ± SD (n = 6). *p <0.05 (one-way ANOVA and Bonferroni post hoc test). (O) Effects of p65 on the activities of different versions of Platr4-Luc reporters. Data are mean ± SD (n = 6). *p <0.05 (t-test). (P) Recruitment of p65 protein to the κB sites (-1066/-1056 bp and -526/-516 bp) of Platr4 promoter in unstimulated or Tnfα-stimulated BMDMs. Data are mean ± SD (n = 3). *p <0.05 (two-way ANOVA and Bonferroni post hoc test). NR, non-binding region.