Fig 1: Summary of temporal expression profiles of key transcription factors in the SC lineage.Sox9 and Sox10 are expressed throughout SC genesis, beginning at E9.0 in migrating NCCs and persisting until P65 in myelinating and non-myelinating SCs in the sciatic nerve. AP2α, Pax3 and Etv5 are also expressed in NCCs, persisting until E12.5 in SCPs with Nfatc4 expression being restricted to NCCs. Egr1 and Sox2 expression is initiated in SCPs at E12.5, persisting until E14.5 in iSCs. At E18.5, AP2α is expressed in the DRG but is undetectable in the nerves while the expression of Jun, Oct6, and Yy1 is initiated and persists till P65. Rare Sox2+Sox10+ positive cells are also detected in the P65 nerve. Egr2 expression is not detected until P7 in myelinating SCs in the sciatic nerve. Post-injury, high upregulation in expression of Sox2, Oct6, and Jun is observed, while distinct nuclear Egr1 expression is also detected. The straight lines represent continued expression of the markers through the different stages, while the dotted lines represent declining or low expression. The asterisk indicates where expression is restricted to the DRG and is undetectable in the nerves at E18.5. Markers expressed in the nerve after an acute injury have been denoted with ‘√’. The green cells represent the developing axon, while the beige cells represent the NCCs at E9.0-E10.5 and the developing SCs at E12.5 to post-natal stages.
Fig 2: Administration of either FK506 or 11R-vivit inhibits NFAT3 nuclear translocation, suppresses FasL expression, attenuates apoptosis and necroptosis in OHCs, and protects against NIHL. (A) Quantification analysis of abnormal OHCs nuclei (abnormal OHCs/total OHCs) in the basal turn of cochlear epithelia 2 h after noise exposure. (B) The auditory threshold shift was measured by ABR test 2 weeks after noise exposure. (C) Quantification analysis of OHCs loss in the entire cochlea 2 weeks after noise exposure. (D) Multiple immunofluorescence staining for FasL/NFAT3/phalloidin/Hoechst33342 was performed on the basal turn of cochlear epithelia immediately after noise exposure. Bottom insets display local magnifications. Semi-quantification analysis of the immunofluorescence intensity reveals that FK506 and 11R-vivit significantly reduced noise-induced FasL expression (E) and NFAT3 nuclear accumulation (F) and in OHCs. N = 5 mice. (G) Western blot analysis of cochlear homogenates 2 h after noise exposure, measuring the expression levels of P-MLKL, MLKL, FasL and cleavage caspase 3. (H) Semi-quantitative analyses of the relative band densities of P-MLKL, MLKL, FasL, and cleavage caspase 3 compared with GAPDH. Scale bar = 10 μm; *, #p < 0.05, **, ##p < 0.01, ***p < 0.001; ns: not significant; N = 5 mice.
Fig 3: The validation of obviously differentially expressed RNAs in the ceRNA network. (A) The ceRNA network of KLRD1, BRAF, and NFATC4 (rectangles, DEmiRNAs; triangles, DEcircRNAs; diamonds, DElncRNAs; Circles, DEmRNAs). (B) The fold changes of KLRD1, BRAF, and NFATC4 were involved in ceRNA regulatory network in the ICP group compared with the control group. (C) The validation on RNA expression in the ICP group (n=3) compared with the control group (n=3) by qRT-PCR. Each dot represents the average value of one sample in three experimental replicates. (D) The protein level of NFATC4 and BRAF in the ICP group compared with the control group. The letters represented the different placenta tissues. (E) Comparison of cytokines levels in placenta tissue between ICP and healthy controls. Cytokines concentrations of TNF-α, IFN-γ and IL-10 were compared between ICP patients (n=7) and healthy controls (n=5). Data were shown as mean ± SD. Differences were analyzed by unpaired t test. All P-values are two-tailed and significantly different when P-value is <0.05. *P<0.05, **P<0.01, ***P<0.001.
Fig 4: Role of hsa-mir-1180-5p in NFATC4 protein expression and function.A: Inhibition of radiation-induced downregulation of NFATC4 protein levels by blocking the effect of hsa-mir-1180-5p. A549 cells were treated with the hsa-mir-1180-5p inhibitor or inhibitor control for 24 h and irradiated with 2 Gy of radiation. Cells were lysed at the indicated time points and NFATC4 expression was quantified by western blotting. Densitometric quantification was performed on samples from three independent experiments (two-sample t-test combined with Bonferroni’s correction; *p< 0.016 compared to ctr-mir-siRNA). B: A colony formation assay was used to determine the effect of the hsa-mir-1180-p5 inhibitor on the plating efficiency and radio-sensitivity of A549 cells (*p< 0.016 compared to ctr mir-inhibitor).
Fig 5: Changes in expression in rats’ myocardium after S-band radiation. (A) The original graphs of strips; (B) the graphs of statistical analysis:(a) JNK-46 kDa; (b) JNK-54 kDa; (c) p-JNK-46 kDa; (d) p-JNK/JNK; (e) HSF1; (f) NFATc4. Significance signs were classified according to p-value: * p < 0.05 and ** p < 0.01.
Supplier Page from Abcam for Anti-NFATC4 antibody