Fig 1: RUNX1/miR-142-3p/DRD2 pathway regulates the NF-κB-dependent inflammatory responses in H9c2 cells with H/R injury. (A) The NF-κB dependent inflammatory response was regulated by overexpression of RUNX1, miR-142-3p mimic and DRD2 in H9c2 cells with H/R injury and detected using western blotting. (B) Bar diagram showing the protein expression levels of RUNX1, DRD2, P65, TNF-α and IL-6 based on (A). n=3. (C) Bar diagram showing the protein expression of functional p-P65 subunit of NF-κB based on (A). n=3. *P<0.05, **P<0.01. H/R, hypoxia/reoxygenation; DRD2, dopamine receptor D2; p-, phosphorylated-; miR, microRNA; Ctrl, control.
Fig 2: Emodin attenuates the expression level of transcription factor RUNX1 induced by either H/R injury or MIRI. (A) RUNX1 gene expression in H9c2 cells with H/R injury or emodin treatment was determined using RT-qPCR. n=5. (B) The mRNA level of RUNX1 in myocardium with MIRI or emodin treatment was measured using RT-qPCR. n=5. (C) RUNX1 protein was detected by western blotting in H9c2 cells with H/R injury or emodin treatment. (D) Summary for the protein expression level of RUNX1 based on (C). n=3. (E) RUNX1 protein expression in myocardium with MIRI or emodin treatment was detected by western blotting. Three bands represent three independent samples. (F) Summary for the protein expression level of RUNX1 based on (E). n=3. **P<0.01. RUNX1, runt-related transcription factor 1; H/R, hypoxia/reoxygenation; MIRI, myocardial ischemia/reperfusion injury; RT-qPCR, reverse transcription-quantitative PCR; Ctrl, control.
Fig 3: RUNX1 expression in ILCs and heterodimerization with NFκB1. (A, B) The frequency of RUNX1+ cells in Lin-(CD45+NK1.1-FcϵR1α- Lin-) cells and Lin+ (CD45+) cell populations. **P<0.001, N=5 per group. (C) Co-localization of p105/50 and RUNX1 in the lung tissue from Alt/Alt treated Nfκb1+/+. Nfκb1-/- mice were used as controls. (D, E) Co-immunoprecipitation of p105/50 and RUNX1. RUNX1 co-precipitated with p105/50 (D) and conversely, p105/50 co-precipitated with RUNX1 (E). IP, immunoprecipitation and IB, immunoblot.
Fig 4: RUNX1 contributes to heart failure in vivo.(A) Morphological changes of mice heart in different groups under TAC-induced; (B) Ultrasound images of heart structure in different groups under TAC-induced mice; (C) Comparisons of LVIDd, LVIDs, EF and FS value among different groups. Asterisks (*) indicate a significant difference.
Fig 5: RUNX1 can activate TGF-β signaling.(A) Luciferase reporter assay showing that RUNX1 can activate TGF-β signaling in HL-1 cells. TGF-β1 and RUNX1 had cooperative effects in regulating TGF-β signaling. (B) Western blot assay showed that phosphorylation changes of TGF-β downstream signal molecules including Smad2/Smad3 in heart tissues of different groups. Asterisks (*) indicate a significant difference.
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