Journal: Nature Communications

Article Title: Epigenetic activation of the FLT3 gene by ZNF384 fusion confers a therapeutic susceptibility in acute lymphoblastic leukemia

doi: 10.1038/s41467-022-33143-w

Figure Lengend Snippet: a FLT3 expression across different B-ALL subtypes in the US cohort of 1988 children and adults (Cohort 1), with the highest expression of FLT3 in ZNF384 -r ( ZNF384 -rearrangement) ALL ( n = 1988). b FLT3 expression in ALL cases ( n = 49) with various ZNF384 fusions in cohort 1. Subgroups were defined by fusion partners of ZNF384 . Other: fusion partners include ARID1B , CLTC , CREBBP , EWSR1 , NIPBL , and SMARCA2 . c FLT3 expression across different B-ALL subtypes in cohort 2 ( n = 377), with the highest expression of FLT3 in ZNF384 -r ALL. d FLT3 expression across ZNF384 fusions ( n = 17) in the Asian cohort (Cohort 2). Subgroups were defined by fusion partners of ZNF384 . Other: fusion partners include USP25 and CREBBP . e FLT3 mutations in ZNF383-r ALL from each ALL cohort. A.U. arbitrary units. Center lines indicate median values of FLT3 expression. Source data are provided as a Source Data file.

Article Snippet: Antibodies against ZNF384 (Abclonal, #A15964, 4 μg), H3K27ac (CST, #8173 S, 4 μg), or H3K4me3 (CST, #9751, 4 μg) were then incubated with permeabilized cells. pAG-MNase enzyme was then added to digest exposed genomic DNA.

Techniques: Expressing

Journal: Nature Communications

Article Title: Epigenetic activation of the FLT3 gene by ZNF384 fusion confers a therapeutic susceptibility in acute lymphoblastic leukemia

doi: 10.1038/s41467-022-33143-w

Figure Lengend Snippet: a , b ZNF384 CUT&RUN (Cleavage Under Targets & Release Using Nuclease) assay using a TCF3-ZNF384 ALL PDX sample (ID TCZ). In total, 10,470 peaks were identified, with 4820 in promoter regions (±2 kb from TSS) and the other 6650 in enhancers. 6750 peaks overlapped with H3K27ac peaks, while 4602 peaks overlapped with H3K4me3. c CUT&RUN and ATAC-seq (Assay for Transposase-Accessible Chromatin using sequencing) of ALL cell lines and primary ALL blast samples with or without ZNF384 -rearrangements. Each track represents a type of assay in a given sample as indicated on the left. As shown in the CUT&RUN tracks, a unique ZNF384 binding site was observed 25 kb upstream of FLT3 with prominent H3K27ac and H3K4me3 marks. This peak also overlapped with an open chromatin region identified by ATAC-seq in EP300-ZNF384 ALL primary samples ( n = 2). In contrast, this region was void of ATAC-seq signals in ALL cell lines or ALL primary samples of other subtypes. Bottom panel: two ZNF384 binding motifs (AAAAAAAA) were identified by footprint analysis using the ZNF384 CUT&RUN data derived from TCF3-ZNF384 ALL PDX cells. d Enhancer activity of z-FLT3 enhancer was confirmed by luciferase assay. A 500 bp segment (hg38, chr13:28,124,365–28,124,868) covered by this z-FLT3 enhancer increased transcription activity by 12.7-fold compared to vector control in JIH5 cells. Enhancer activity was normalized to empty vector control. Data are shown as mean values ± SEM of three biological replicates (center of the error bar) and the results are representative of three independent experiments. Source data are provided as a Source Data file.

Article Snippet: Antibodies against ZNF384 (Abclonal, #A15964, 4 μg), H3K27ac (CST, #8173 S, 4 μg), or H3K4me3 (CST, #9751, 4 μg) were then incubated with permeabilized cells. pAG-MNase enzyme was then added to digest exposed genomic DNA.

Techniques: Nuclease Assay, Sequencing, Binding Assay, Derivative Assay, Activity Assay, Luciferase, Plasmid Preparation, Control

Journal: Nature Communications

Article Title: Epigenetic activation of the FLT3 gene by ZNF384 fusion confers a therapeutic susceptibility in acute lymphoblastic leukemia

doi: 10.1038/s41467-022-33143-w

Figure Lengend Snippet: ChIA-PET (chromatin interaction analysis with paired-end tags) (CTCF and RNAP II) was performed in both JIH5 cells and an ALL PDX sample harboring the TCF3-ZNF384 fusion. As shown in the top four panels, DNA looping mediated by CTCF and RNAP II was detected between ZNF384-binding site (“z-FLT3 enhancer”) and FLT3 promoter, indicated by shaded box. In addition, we also performed ZNF384 ChIA-PET in both JIH5 cells and TCF3-ZNF384 ALL PDX sample to map ZNF384-mediated chromatin interactions, in comparison to lymphoblastoid cell line GM12878 which does not harbor ZNF384 fusion gene. Red curves indicated chromatin interactions called using ChIA-PIPE (paired-end tag reads ≥2) and were highlighted in red in Supplementary Data and . Additionally, the bottom panels show protein binding sites (ZNF384 and RNAP II) identified from inter-ligation and self-ligation reads of ChIA-PET using MACS2 calling algorithm. Overlap of ZNF384 binding sites and chromatin looping anchors suggests enhancer-promoter interaction specifically mediated by ZNF384 (indicated by green and red arrows).

Article Snippet: Antibodies against ZNF384 (Abclonal, #A15964, 4 μg), H3K27ac (CST, #8173 S, 4 μg), or H3K4me3 (CST, #9751, 4 μg) were then incubated with permeabilized cells. pAG-MNase enzyme was then added to digest exposed genomic DNA.

Techniques: ChIA Pet Assay, Binding Assay, Comparison, Paired-end Tag, Protein Binding, Ligation

Journal: Nature Communications

Article Title: Epigenetic activation of the FLT3 gene by ZNF384 fusion confers a therapeutic susceptibility in acute lymphoblastic leukemia

doi: 10.1038/s41467-022-33143-w

Figure Lengend Snippet: a In vitro sensitivity of a panel of ALL cell lines as well as PDX-derived ALL cell with TCF3-ZNF384 fusion to gilteritinib was determined using MTT assay. Data are shown as mean % viability relative to vehicle ± SEM of three biological replicates (center of the error bar) and results are representative of three independent experiments. b Ex vivo sensitivity of a panel of 47 primary ALL cases including three with EP300-ZNF384 fusion as well as those with ETV6-RUNX1 , DUX4 -r, BCR-ABL1 , hyperdiploidy, and T-ALL. Box plots show summary of data in terms of minimum, maximum, median, and first and third quartiles. Each data point represents two technical replicate. c , d Expression of EPZ ( EP300 - ZNF384 ) fusion gene was downregulated by CRISPR Cas9 editing in JIH5 cells, which led to decreased expression of FLT3 and drug sensitivity to gilteritinib. Data are shown as mean values ± SEM of three biological replicates (center of the error bar) and results are representative of three independent experiments. P values ( P = 0.006) were estimated using two-sided t test. e – h Gilteritinib efficacy was evaluated in vivo using xenograft models. e , f Show leukemia progression, and survival in mice transplanted with JIH5 cells. g , h Describe results of mice transplanted with PDX-derived ALL cells with TCF3-ZNF384 fusion. Gilteritinib was given daily at a dosage of 10 mg/kg, and therapy started three days following leukemia engraftment. Leukemia burden was monitored weekly, and P values ( P < 0.005) were estimated using a two-sided analysis of deviance based on mixed effect models with cubic splines. Leukemia-free survival was plotted as Kaplan–Meier curves and P values were estimated using two-sided log-rank test ( P = 0.0027 for f , P = 0.0026 for h ). Source data are provided as a Source Data file.

Article Snippet: Antibodies against ZNF384 (Abclonal, #A15964, 4 μg), H3K27ac (CST, #8173 S, 4 μg), or H3K4me3 (CST, #9751, 4 μg) were then incubated with permeabilized cells. pAG-MNase enzyme was then added to digest exposed genomic DNA.

Techniques: In Vitro, Derivative Assay, MTT Assay, Ex Vivo, Expressing, CRISPR, In Vivo

Journal: Computational and Structural Biotechnology Journal

Article Title: Bulk RNA-seq and scRNA-seq analysis reveal an activation of immune response and compromise of secretory function in major salivary glands of obese mice

doi: 10.1016/j.csbj.2022.11.054

Figure Lengend Snippet: The levels of immune response related proteins were elevated in the SGs of DIO mice. (A-C) Immunohistochemical analysis of CD4 (A), CD8 (B) and CD79A (C) in the SMG, SLG and PG from the NC and DIO mice (week-35), respectively. Arrow points to the representative region with elevated protein positive signals in the DIO mice. (D) Western blot analysis of CD4, CD79A, CD8A, MUC20, MUC13 and α-Amylase in the SGs of three NC mice and three DIO mice (week-35), respectively. β-Actin: loading control.

Article Snippet: Antibodies against CD4 (ER1706-80, HuaBio, Hangzhou), CD8A (0108–7, HuaBio, Hangzhou), CD79A (EM1902-29, HuaBio, Hangzhou), MUC13 (ER1913-37, HuaBio, Hangzhou), MUC20 (A15968, ABclonal,Wuhan), α-Amylase (3796,Cell Signaling Technology, America) andβ-Actin (AC026, ABclonal, Wuhan) were purchased.

Techniques: Immunohistochemical staining, Western Blot

Journal: Cell Research

Article Title: In vivo self-assembled small RNAs as a new generation of RNAi therapeutics

doi: 10.1038/s41422-021-00491-z

Figure Lengend Snippet: a Flow chart of the experimental design. Male C57BL/6J mice at 3 weeks of age were placed on a HFD for 12 weeks. Mice rapidly gained weight and became obese. Mice were then maintained on a HFD and treated with PBS or 5 mg/kg CMV-scrR, CMV-siR P or CMV-RVG-siR P circuit through tail vein injection for a total of 12 times over 24 days. Body weights were monitored during treatment. After treatment, mice were divided into several groups and subjected to the evaluation of fat mass, energy expenditure, leptin sensitivity and glucose homoeostasis. b Body weight curves ( n = 14 in each group). c Weights of epididymal fat pads ( n = 14 in each group). d – k Energy expenditure parameters, including oxygen consumption (VO 2 ), respiratory exchange ratio (RER), total activity and heat production, were monitored ( n = 3 in each group). l , m Weight loss and food intake inhibition in response to leptin. Male mice were injected with leptin (0.5 μg/g body weight every 12 h) for the indicated periods. Food intake and body weight were monitored 2 days prior to the start of injection and normalized to 100% for day 0 values ( n = 6 in each group). n Basal serum leptin levels ( n = 6 in each group). o Western blot analysis of PTP1B in the hypothalamus. Mice were injected with or without leptin (to stimulate leptin signalling), and hypothalamus was collected for immunoblot analysis of PTP1B ( n = 3 in each group). SSG (200 mg/kg i.p. dose) serves as a control. Shown are representative western blots. p Mouse GTT results ( n = 8 in each group). q Mouse ITT results. Blood glucose values are expressed as the percentage of the initial concentration ( n = 8 in each group). r Western blot analysis of PTP1B and tyrosine phosphorylation of insulin receptors in the liver. Mice were injected with or without insulin (to stimulate insulin signalling), and liver was collected for immunoblot analysis with antibodies against PTP1B, p-IR (Tyr1162/Tyr1163) or total IR ( n = 3 in each group). SSG (200 mg/kg i.p. dose) serves as a control. Shown are representative western blots. Values are presented as the means ± SEM. Significance was determined using one-way ANOVA followed by Dunnett’s multiple comparison in c , e , g , i , k , n , and using two-way ANOVA followed by Dunnett’s multiple comparison in b , l , m , p , q . * P < 0.05; ** P < 0.01; *** P < 0.005; ns, not significant.

Article Snippet: Antibodies used were listed as below: anti-EGFR (A2B1) (18986-1-AP), anti-KRAS (12063-1-AP) and anti-p-AKT (Ser473) (66444-1-Ig) antibodies were purchased from Proteintech (IL, USA), and the anti-KRAS antibody (12063-1-AP, Proteintech) recognized total KRAS (wild-type KRAS and mutant KRAS) rather than KRAS G12D ; anti-GAPDH (G-9) (sc-365062) antibody was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA); anti-TNC (ab108930) and anti-IR (ab131238) antibodies were purchased from Abcam (Cambridge, MA, USA); anti-Flag antibody (MA1-91878) was purchased from Thermo Fisher (San Jose, CA, USA); anti-PTP1B (A1590) antibody was purchased from ABclonal (Cambridge, MA, USA); anti-p-Erk1/2 (Thr202/Tyr204) (4376) antibody was purchased from Cell Signalling Technology (Danvers, MA, USA); and anti-p-IR/IGF1R (Tyr1158/Tyr1162/Tyr1163) (P06213) antibody was purchased from Millipore (Bedford, MA, USA).

Techniques: Injection, Activity Assay, Inhibition, Western Blot, Control, Concentration Assay, Phospho-proteomics, Comparison

Journal: CNS Neuroscience & Therapeutics

Article Title: Cdk5 phosphorylation‐dependent C9orf72 degradation promotes neuronal death in Parkinson's disease models

doi: 10.1111/cns.14319

Figure Lengend Snippet: Chromosome 9 open reading frame 72 (C9orf72) expression is decreased in Parkinson's disease (PD) animal models. (A) Western blotting of C9orf72, p62, α‐synuclein, and LC3 in MPTP‐induced PD mice. (B) Statistical analyses of the relative content of C9orf72 in MPTP‐induced PD mice. (C) Statistical analysis of the relative content of p62 in MPTP‐induced PD mice. (D) Statistical analysis of the relative content of α‐synuclein in MPTP‐induced PD mice. (E) Statistical analysis of the relative content of LC3 in MPTP‐induced PD mice, n = 3/group. * p < 0.05. (F) Western blotting of C9orf72, p62, α‐synuclein, and LC3 in α‐synuclein A53T‐induced genetic PD mice. (G) Statistical analysis of the relative content of α‐synuclein A53T‐induced C9orf72. (H) Statistical analysis of the relative content of α‐synuclein A53T‐induced p62. (I) Statistical analysis of the relative content of α‐synuclein A53T‐induced α‐synuclein. (J) Statistical analysis of the relative content of α‐synuclein A53T‐induced LC3, n = 3/group, ** p < 0.01, * p < 0.05. (K) Western blotting of C9orf72, p62, α‐synuclein, and LC3 in preclinical PFF‐induced PD mice. (L) Statistical analysis of the relative content of C9orf72 in preclinical PFF‐induced PD mice. (M) Statistical analysis of the relative content of p62 in preclinical PFF‐induced PD mice. (N) Statistical analysis of the relative content of α‐synuclein in preclinical PFF‐induced PD mice. (O) Statistical analysis of the relative content of LC3, n = 3/group, ** p < 0.01, * p < 0.05. Data are presented as the mean ± SEM.

Article Snippet: Rabbit polyclonal anti‐C9orf72 (WB‐1: 800, A15970) was purchased from Abclonal.

Techniques: Expressing, Western Blot

Journal: CNS Neuroscience & Therapeutics

Article Title: Cdk5 phosphorylation‐dependent C9orf72 degradation promotes neuronal death in Parkinson's disease models

doi: 10.1111/cns.14319

Figure Lengend Snippet: Chromosome 9 open reading frame 72 (C9orf72) expression was decreased in Parkinson's disease (PD) cellular models. (A) Western blotting of C9orf72, p62, α‐synuclein, and LC3 in MPP + ‐induced neurons. (B) Statistical analysis of the relative content of C9orf72 in MPP + ‐induced neurons. (C) Statistical analysis of the relative content of p62 in MPP + ‐induced neurons. (D) Statistical analysis of the relative content of α‐synuclein in MPP + ‐induced neurons. (E) Statistical analysis of the relative content of LC3 in MPP + ‐induced neurons. (F) Western blotting of C9orf72, p62, α‐synuclein, and LC3 in MPP + ‐induced SH‐SY5Y cells. (G) Statistical analysis of the relative content of C9orf72 in MPP + ‐induced SH‐SY5Y cells. (H) Statistical analysis of the relative content of p62 in MPP + ‐induced SH‐SY5Y cells. (I) Statistical analysis of the relative content of α‐synuclein in MPP + ‐induced SH‐SY5Y cells. (J) Statistical analysis of the relative content of LC3 in MPP + ‐induced SH‐SY5Y cells. (K) Western blotting of C9orf72, p62, α‐synuclein, and LC3 in an A53T‐induced SY5Y genetic model. (L) Statistical analysis of the relative content of C9orf72 in an A53T‐induced SY5Y genetic model. (M) Statistical analysis of the relative content of p62 in an A53T‐induced SY5Y genetic model. (N) Statistical analysis of the relative content of α‐synuclein in an A53T‐induced SY5Y genetic model. (O) Statistical analysis of the relative content of LC3 in an A53T‐induced SY5Y genetic model. (P) Western blotting of C9orf72, p62, α‐synuclein, and LC3 in a PFF‐induced neurons preclinical model. (Q) Statistical analysis of the relative content of C9orf72 in a PFF‐induced neurons preclinical model. (R) Statistical analysis of the relative content of p62 in a PFF‐induced neurons preclinical model. (S) Statistical analysis of the relative content of α‐synuclein in a PFF‐induced neurons preclinical model. (T) Statistical analysis of the relative content of LC3 in a PFF‐induced neurons preclinical model. n = 3/group, ** p < 0.01, * p < 0.05. Data are presented as the mean ± SEM.

Article Snippet: Rabbit polyclonal anti‐C9orf72 (WB‐1: 800, A15970) was purchased from Abclonal.

Techniques: Expressing, Western Blot

Journal: CNS Neuroscience & Therapeutics

Article Title: Cdk5 phosphorylation‐dependent C9orf72 degradation promotes neuronal death in Parkinson's disease models

doi: 10.1111/cns.14319

Figure Lengend Snippet: Knockdown of Chromosome 9 open reading frame 72 (C9orf72) aggravates autophagy and the overexpression of C9orf72 rescues autophagy. (A) Western blotting of p62, α‐synuclein, and LC3 in C9orf72‐overexpressing SY5Y cells induced by MPP + . (B) Statistical analysis of the relative content of p62 in C9orf72‐overexpressing SY5Y cells induced by MPP + . (C) Statistical analysis of the relative content of α‐synuclein in C9orf72‐overexpressing SY5Y cells induced by MPP + . (D) Western blotting of LC3 in C9orf72‐overexpressing SY5Y cells induced by MPP + . (E) Western blotting of C9orf72, p62, α‐synuclein, and LC3 in C9orf72‐knockdown mice. (F) Statistical analysis of the relative content of C9orf72 in C9orf72‐knockdown mice. (G) Statistical analysis of the relative content of p62 in C9orf72‐knockdown mice. (H) Western blotting of α‐synuclein in C9orf72‐knockdown mice. (I) Statistical analysis of the relative content of LC3 in C9orf72‐knockdown mice. n = 3/group, *** p < 0.001, ** p < 0.01, * p < 0.05. (J) Immunofluorescence staining of TH enzyme in C9orf72‐knockdown mice. (K) TH+ neuron count and statistical analysis in C9orf72‐knockdown mice. n = 3/group. *** p < 0.001, ** p < 0.01, * p < 0.05. Data are presented as the mean ± SEM.

Article Snippet: Rabbit polyclonal anti‐C9orf72 (WB‐1: 800, A15970) was purchased from Abclonal.

Techniques: Over Expression, Western Blot, Immunofluorescence, Staining

Journal: CNS Neuroscience & Therapeutics

Article Title: Cdk5 phosphorylation‐dependent C9orf72 degradation promotes neuronal death in Parkinson's disease models

doi: 10.1111/cns.14319

Figure Lengend Snippet: Chromosome 9 open reading frame 72 (C9orf72) protein is degraded by the ubiquitination degradation pathway. (A) 3‐MA and MG132 were used to intervene in the autophagy‐lysosomal pathway and the ubiquitin‐proteasome pathway, respectively, in order to detect the degradation of C9orf72 protein after MPP + treatment of primary neurons for 24 h. (B) Statistical analysis of the relative content of C9orf72. n = 3, * p < 0.05. Data are presented as the mean ± SEM.

Article Snippet: Rabbit polyclonal anti‐C9orf72 (WB‐1: 800, A15970) was purchased from Abclonal.

Techniques:

Journal: CNS Neuroscience & Therapeutics

Article Title: Cdk5 phosphorylation‐dependent C9orf72 degradation promotes neuronal death in Parkinson's disease models

doi: 10.1111/cns.14319

Figure Lengend Snippet: Cdk5 is involved in the degradation of Chromosome 9 open reading frame 72 (C9orf72). (A) GPS3.0 predicts that the S9 site of the C9orf72 protein can be phosphorylated by Cdk5. (B) Scansite predicted that Cdk5 phosphorylates the S9 site of the C9orf72 protein. (C) PhosphoSitePlus revealed that the cC9orf72 S9 site could be phosphorylated in the mass spectrometry results of other scholars. (D) DNAMAN alignment of the known amino acid conservation at the S9‐site vicinity of human and mouse C9orf72 proteins. (E) C9orf72 interacted with Cdk5. (F) HEK293 cells were co‐transfected with HA‐Cdk5/Myc‐p35, Flag‐C9orf72 WT plasmid, or Flag‐C9orf72 S9A plasmid, and Phos S/TP antibody was used to detect the phosphorylation results. (G) After 24 h of HEK293 cells' transfection with HA‐Cdk5/Myc‐p35, wild‐type Flag‐C9orf72 WT, or mutant Flag‐C9orf72 S9A plasmid. Western blotting to detect the expression of C9orf72 protein. (H) Detection of the phosphorylation level of C9orf72 protein in MPTP‐induced mice. (I) Detection of the phosphorylation level of C9orf72 protein in MPP + ‐induced primary neurons.

Article Snippet: Rabbit polyclonal anti‐C9orf72 (WB‐1: 800, A15970) was purchased from Abclonal.

Techniques: Mass Spectrometry, Transfection, Plasmid Preparation, Mutagenesis, Western Blot, Expressing

Journal: CNS Neuroscience & Therapeutics

Article Title: Cdk5 phosphorylation‐dependent C9orf72 degradation promotes neuronal death in Parkinson's disease models

doi: 10.1111/cns.14319

Figure Lengend Snippet: Cdk5 inhibitors inhibited the phosphorylation degradation of Chromosome 9 open reading frame 72 (C9orf72). (A) Western blotting of Cdk5 inhibitor ROSCOVITINE on the expression of C9orf72, p62, α‐synuclein, and LC3 in MPP + ‐induced neurons. (B) Statistical analysis of the relative content of C9orf72 in MPP + ‐induced neurons inhibited by ROSCOVITINE. (C) Statistical analysis of the relative content of p62 in MPP + ‐induced neurons inhibited by ROSCOVITINE. (D) Statistical analysis of the relative content of α‐synuclein in MPP + ‐induced neurons inhibited by ROSCOVITINE. (E) Statistical analysis of the relative content of LC3 in MPP + ‐induced neurons inhibited by ROSCOVITINE. (F) Western blotting of the Cdk5 inhibitor ROSCOVITINE on the expression of C9orf72, p62, α‐synuclein, and LC3 in MPP + ‐induced SY5Y cells. (G) Statistical analysis of the relative content of C9orf72 in MPP + ‐induced SY5Y cells inhibited by ROSCOVITINE. (H) Statistical analysis of the relative content of p62 in MPP + ‐induced SY5Y cells inhibited by ROSCOVITINE. (I) Statistical analysis of the relative content of α‐synuclein in MPP + ‐induced SY5Y cells inhibited by ROSCOVITINE. (J) Statistical analysis of the relative content of LC3 in MPP + ‐induced SY5Y cells inhibited by ROSCOVITINE. (K) Western blotting of the Cdk5 inhibitor ROSCOVITINE on the expression of C9orf72, p62, α‐synuclein, and LC3 in PFF‐induced neurons. (L) Statistical analysis of the relative content of C9orf72 in PFF‐induced neurons inhibited by ROSCOVITINE. (M) Statistical analysis of the relative content of p62 in PFF‐induced neurons inhibited by ROSCOVITINE. (N) Statistical analysis of the relative content of α‐synuclein in PFF‐induced neurons inhibited by ROSCOVITINE. (O) Statistical analysis of the relative content of LC3 in PFF‐induced neurons inhibited by ROSCOVITINE. n = 3/group, ** p < 0.01, * p < 0.05. Data are presented as the mean ± SEM.

Article Snippet: Rabbit polyclonal anti‐C9orf72 (WB‐1: 800, A15970) was purchased from Abclonal.

Techniques: Western Blot, Expressing

Journal: CNS Neuroscience & Therapeutics

Article Title: Cdk5 phosphorylation‐dependent C9orf72 degradation promotes neuronal death in Parkinson's disease models

doi: 10.1111/cns.14319

Figure Lengend Snippet: Myr‐Chromosome 9 open reading frame 72 (C9orf72) peptide rescues neuronal loss and motor dysfunction. A The effect of different concentrations of Myr‐C9orf72 peptide on the viability of MPP + ‐treated primary neurons. n = 3. *** p < 0.001, ** p < 0.01. B Western blotting of C9orf72, p62, α‐synuclein, and LC3. C Statistical analysis of the relative content of C9orf72. D Statistical analysis of the relative content of p62. E Western blotting of α‐synuclein. F Statistical analysis of the relative content of LC3. n = 3/group, ** p < 0.01, * p < 0.05. G Immunofluorescence staining of TH. H TH+ neuron count and statistical analysis. n = 3/group. *** p < 0.001, * p < 0.05. I Rotarod test data after the experiment. J Open field data after the experiment. n = 12/group, compared with the control group, *** p <0.001, ** p <0.01. Data are presented as the mean ± SEM.

Article Snippet: Rabbit polyclonal anti‐C9orf72 (WB‐1: 800, A15970) was purchased from Abclonal.

Techniques: Western Blot, Immunofluorescence, Staining

Journal: CNS Neuroscience & Therapeutics

Article Title: Cdk5 phosphorylation‐dependent C9orf72 degradation promotes neuronal death in Parkinson's disease models

doi: 10.1111/cns.14319

Figure Lengend Snippet: The theoretical scheme of Chromosome 9 open reading frame 72 (C9orf72) protein involved in neuronal death in Parkinson's disease (PD). During PD, Cdk5 is activated in dopaminergic neurons of the substantia nigra, and the activated protein kinase Cdk5 phosphorylates the S9 site of the C9orf72 protein. Phosphorylation at the S9 site mediates C9orf72 protein degradation through the ubiquitin‐proteasome pathway. The degradation and loss of function of the C9orf72 protein led to autophagy impairment in neurons. Proteins such as p62 and α‐synuclein, which cannot be effectively cleared by autophagy, accumulate abnormally, and their neurotoxicity causes the death of dopaminergic neurons.

Article Snippet: Rabbit polyclonal anti‐C9orf72 (WB‐1: 800, A15970) was purchased from Abclonal.

Techniques: