Fig 1: Drp1/Fis1 axis accelerates mitochondrial dysfunction in the cerebral cortex of CKD mice. A Heat map of expression of upregulated genes in RNA-Seq. B Venn map of intersection between upregulated genes in RNA-Seq and “Renal Failure”-related genes in GeneCards and DisGeNET. C The genes interacted with Fis1 predicted by STRING. The redder the circle where the gene located indicates the higher combined_score. Drp1 (DNM1L) has the highest combined_score. D FIS1 and Drp1 expression in RNA-Seq cerebral cortex sample of normal mice (blue, n = 3) and CKD mice (red, n = 3). E mRNA levels of Fis1 and Drp1 in the cerebral cortex of CKD mice determined by RT-qPCR. F Expression of Fis1 and Drp1 proteins in the cerebral cortex of CKD mice detected by immunohistochemical staining. G–J MDA level (G), SOD activity (H), ATP level (I), and ROS level (J) in the cerebral cortex of CKD mice. K The morphology of mitochondria in cerebral cortex of CKD mice observed under the TEM (scale bar = 1 μm) and mean mitochondria size. N = 6. *p < 0.05
Fig 2: CKD-induced hypercalcemia mediates Drp1/Fis1 to promote mitochondrial fragmentation and aggravate nerve damage. CKD mice were treated with calcitonin alone or combined with overexpressed Drp1 (n = 6). A Ca2+ content in serum of CKD mice. B Ca2+ content in prefrontal cortical tissues of CKD mice. C Expression of Drp1 and Fis1 proteins in prefrontal cortical tissues of CKD mice detected by immunohistochemical staining. D The escape latency, the number of times entering the target quadrant, the time spent in the platform quadrant, and the total distance of action of CKD mice by Morris water maze test. E–H MDA level (E), SOD activity (F), ATP level (G), and ROS level (H) in the prefrontal cortical tissues of CKD mice. I The mean mitochondria size in cerebral cortex of CKD mice. J Apoptosis in prefrontal cortical tissues of CKD mice detected by TUNEL staining. K Proportion of NeuN-positive cells in prefrontal cortical tissues of CKD mice detected by immunofluorescence staining. N = 6. *p < 0.05
Fig 3: miR‐181a/b‐1 downregulation leads to an increase of mitochondrial proteins A–EImmunofluorescence analysis showed a decrease of Citrate Synthase (CS) staining, a mitochondrial marker, in the OS of P347S retinas versus WT (A) at p12 (B) and p30 (D). The analysis highlights an amelioration of CS staining in P347S/miR‐181a/b+/− versus P347S eyes at both p12 (C) and p30 (E) (Red triangles). (A′–E′) show higher magnification of (A–E). Scale bars 25 μm.F, GWB analysis reveals decreased levels of OXPHOS, representative of Mitochondrial Respiratory Chain complexes, and CS proteins in the eyes of P347S with respect to WT, and partial rescue of these parameters in P347S/miR‐181a/b+/− versus P347S PR at p30 [quantified in (G)]. Data are normalized to p115. N = 3 eyes/genotype.H, IWB analysis of key proteins involved in mitochondrial fission/fusion pathway (Fis1 Opa1 and Mfn1/2 proteins) in the eyes of WT, P347S and in P347S/miR‐181a/b+/− at p30 [quantified in (I)]. Data are normalized to Gapdh. N ≥ 4 eye/genotype. Data information: Data are presented as mean of Fold Change ± SEM. Student's t‐test, unpaired. Please note that all compared bands from WT, P347S and P347S/miR‐181a/b+/− samples are from the same blots, which were cropped and shown organized in the panel for the sake of data presentation clarity (see source data). Source data are available online for this figure.
Fig 4: Schematic representation of the role of JTE-013 in pulmonary fibrosis. JTE-013 or S1PR2 knockdown inhibited the binding of S1P and S1PR2 to regulate the RHOA/YAP pathway. This further affected the downstream CTGF/CYR61 expression and mitochondrial dynamics, promoted mitochondrial fusion and the phenotypic transition of MFN, inhibited mitochondrial fission, down-regulated the phosphorylation level of Fis1 and Drp1, and reduced the production of mitochondrial ROS and ROS, thereby alleviating pulmonary fibrosis.
Fig 5: JTE-013 inhibited the expression of mitochondrial fusion/fission protein and fibrosis marker proteins in lung tissues. (A): Immunofluorescence staining was used to detect the colocalization of Drp1 and Tom20 (scale bar = 200 μm). (B): TUNEL staining was used to detect apoptosis in lung tissue. (C): Western blot was used to detect the expression of Fis1, OPA1, and MFN1/2 and the phosphorylation and transformation of Drp1. (D): Immunohistochemical staining was used to detect the expression of α-SMA, COL1A1, and MMP-9 and their OD quantitative analysis chart (scale bar = 50 μm). (E): Western blot was used to detect the expression of α-SMA, COL1A1, and MMP-9 (** p < 0.01 vs. control group, ## p < 0.01 vs. BLM group, && p < 0.01 vs. BLM + JTE-013 group). Original Western blot available in Supplementary Materials.
Supplier Page from Abcam for Anti-TTC11/FIS1 antibody