Figure 7.
Inflammatory cytokines may upregulate hepcidin in PV. (A) Mean-difference plot showing the average log expression of each gene (x-axis) and their log-fold change between PV and control liver samples (y-axis). The DEGs are highlighted with points in red and blue indicating upregulated and downregulated genes, respectively (adjusted P value <.05). (B) Heatmap of the expression of all DEGs with hierarchical clustering in which expression values are standardized to have mean of 0 and standard deviation of 1 for each gene. (C) Bar chart depicting GO biological processes, MSigDB hallmark gene sets, or KEGG pathways relating to JAK-STAT signaling, inflammatory response, IL-6 responses, or cytokine-cytokine receptor interactions that are associated with upregulated genes in PV liver samples vs control. x-axis represents statistical significance of the enrichment, increasing from left to right. The red dotted line represents P = .05. (D) HAMP mRNA expression of HepG2 cells cultured in media supplemented with 2% plasma from HC donors or patients with PV; HC, n = 4; PV, n = 3. (E) Mouse serum IL-6; control, n = 8; PV, n = 10. (F-H) Liver Saa1 (F), Fga (G), and Hamp1 (H) relative to Hprt; control + anti–immunoglobulin G (IgG), n = 14; control + anti–IL-6, n = 9; PV + anti-IgG, n = 10; and PV + anti–IL-6, n = 13. (I) SMAD7 and FGA mRNA expression of HepG2 cells cultured in media supplemented with 2% plasma from HC donors or patients with PV; n = 4. (J-K) HAMP mRNA expression of HepG2 (J) or Huh7 (K) cells cultured in media supplemented with 10 ng/mL recombinant human IL-6–family cytokines; n = 3. The red line in panel J indicates HAMP expression in the absence of additional cytokines. (L) HAMP mRNA expression of HepG2 cells cultured in media supplemented with 2% plasma from HC donors or patients with PV with the addition of anti-GP130 antibodies or vehicle control (phosphate-buffered saline); n = 4. Unpaired 2-tailed t test with Welch correction for panels D-E, Kruskal-Wallis test for panels F-H, or two-way ANOVA with Šídák correction for multiple comparisons for panels I,L. ∗P < .05; ∗∗P < .01; ∗∗∗P < .001; ∗∗∗∗P < .0001. GO, gene ontology; OSM, oncostatin M; LIF, leukemia inhibitory factor; CT-1, cardiotrophin 1; CNTF, ciliary neurotrophic factor; CLCF1, cardiotrophin-like cytokine factor.

Inflammatory cytokines may upregulate hepcidin in PV. (A) Mean-difference plot showing the average log expression of each gene (x-axis) and their log-fold change between PV and control liver samples (y-axis). The DEGs are highlighted with points in red and blue indicating upregulated and downregulated genes, respectively (adjusted P value <.05). (B) Heatmap of the expression of all DEGs with hierarchical clustering in which expression values are standardized to have mean of 0 and standard deviation of 1 for each gene. (C) Bar chart depicting GO biological processes, MSigDB hallmark gene sets, or KEGG pathways relating to JAK-STAT signaling, inflammatory response, IL-6 responses, or cytokine-cytokine receptor interactions that are associated with upregulated genes in PV liver samples vs control. x-axis represents statistical significance of the enrichment, increasing from left to right. The red dotted line represents P = .05. (D) HAMP mRNA expression of HepG2 cells cultured in media supplemented with 2% plasma from HC donors or patients with PV; HC, n = 4; PV, n = 3. (E) Mouse serum IL-6; control, n = 8; PV, n = 10. (F-H) Liver Saa1 (F), Fga (G), and Hamp1 (H) relative to Hprt; control + anti–immunoglobulin G (IgG), n = 14; control + anti–IL-6, n = 9; PV + anti-IgG, n = 10; and PV + anti–IL-6, n = 13. (I) SMAD7 and FGA mRNA expression of HepG2 cells cultured in media supplemented with 2% plasma from HC donors or patients with PV; n = 4. (J-K) HAMP mRNA expression of HepG2 (J) or Huh7 (K) cells cultured in media supplemented with 10 ng/mL recombinant human IL-6–family cytokines; n = 3. The red line in panel J indicates HAMP expression in the absence of additional cytokines. (L) HAMP mRNA expression of HepG2 cells cultured in media supplemented with 2% plasma from HC donors or patients with PV with the addition of anti-GP130 antibodies or vehicle control (phosphate-buffered saline); n = 4. Unpaired 2-tailed t test with Welch correction for panels D-E, Kruskal-Wallis test for panels F-H, or two-way ANOVA with Šídák correction for multiple comparisons for panels I,L. ∗P < .05; ∗∗P < .01; ∗∗∗P < .001; ∗∗∗∗P < .0001. GO, gene ontology; OSM, oncostatin M; LIF, leukemia inhibitory factor; CT-1, cardiotrophin 1; CNTF, ciliary neurotrophic factor; CLCF1, cardiotrophin-like cytokine factor.

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