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. 2023 May 18;16(3):411–429. doi: 10.1016/j.jcmgh.2023.05.002

Figure 7.

Figure 7

TGF-β signaling and its downstream molecules are up-regulated in DSS-induced colitis Nox4-/-mice. (A) Local Fisher discriminant analysis plot for principal component analysis. Two-dimensional scores are readily distinguished between W/DSS WT (green) and W/DSS Nox4-/- (red) samples. The shared areas indicate 95% CIs. (B) Total colonic cells sorted from W/DSS WT or W/DSS Nox4-/- were subject to RNA-seq. The heatmap of RNA-seq data showing up-regulated (purple) and down-regulated (yellow) genes. (C) Volcano plot identifying significantly up-regulated genes in W/DSS WT (white) and W/DSS Nox4-/- (red). CIBERSORT analysis of WT and Nox4-/- colon tissues. The relative proportions of (D) 11 different immune cell types and (E) 7 T-cell lineage cell types are deconvoluted from the RNA-seq data using CIBERSORT. (F) Bubble plot showing significantly enriched KEGG pathways in the W/DSS WT and W/DSS Nox4-/- DEGs. The bubble colors indicate P values, and the bubble size represents the expression level. (G) Heatmap of the expression of TGF-β–related genes from W/DSS Nox4-/- DEGs. (H) Immunoblotting of TGF-β and TGFβR1 protein levels from W/DSS WT or W/DSS Nox4-/- distal colon tissue extracts. β-actin was used as an internal control. The band size was quantified by ImageJ. (I) Flow cytometric plots showing the expression of RORγT on CD4+ T cells from W/DSS WT and W/DSS Nox4-/- colon tissues. Data are expressed as means ± SD. ∗∗∗∗P < .001 compared with W/DSS WT. JAK-STAT, Janus kinase–signal transducer and activator of transcription pathway; MAPK, mitogen-activated protein kinase; mTOR, Mammalian target of rapamycin; NK, natural killer cell; p-val, P value; TCR, T cell receptor.