Journal: Nature

Article Title: DNA damage burden causes selective CUX2 neuron loss in neuroinflammation

doi: 10.1038/s41586-026-10310-3

Figure Lengend Snippet: a , Classification of 698 DDR genes according to repair pathway (left) or other biological process mediating DDR (right). b , Classification of 159 DDR DEG in MS L2/3ENs according to DNA repair pathway ( n = 12 MS, n = 9 CTRL). c , Geneset enrichment plot for DDR genes ( n = 12 MS, n = 9 CTRL; FDR 0.05, log2 fold change 0.1). d , Box plot of DDR DEG across cell subtypes in human MS ( n = 12 MS, n = 9 CTRL). Plot depicts median and interquartile range (IQR) of DEG calculated after downsampling (100 DEG analyses per cell cluster). e , Pseudotime trajectory ( e ) and bean plots ( f ) of top down (blue) and upregulated (red) DDR DEG in L2/3ENs ( n = 12 MS, n = 9 CTRL). g , Bean expression plots of base excision repair (BER), double-strand break repair (DSBR), single-strand break repair (SSBR) and nucleotide excision repair (NER) factors in MS L2/3ENs ( n = 12 MS, n = 9 CTRL). FAP = Fanconi Anaemia pathway, MMR = mismatch repair, NHEJ = non-homologous end joining, HDR = homology directed repair.

Article Snippet: Non-blunt NHEJ reporter plasmids were a gift from E. Rajendra (Artios Pharma) and prepared by I-SceI digestion and purification as described previously .

Techniques: Expressing, Non-Homologous End Joining

Journal: Nature

Article Title: DNA damage burden causes selective CUX2 neuron loss in neuroinflammation

doi: 10.1038/s41586-026-10310-3

Figure Lengend Snippet: a , Schematic of the alkaline comet assay(top) and representative SYBR-Green-stained images (middle), assessing DNA damage (D) and recovery (R) over 0–4 h in SH-SY5Y cells with GFP, CUX2 or ATF4 overexpression (OE), either untreated (UT) or acutely treated with TBHP (200 µM, 1 h). Quantification shows the recovery-to-damage (R/D) tail moment ratios. n = 3 (ATF4-OE groups and CUX2-OE UT) and n = 4 (other groups). b , NGN2 -iPS cell differentiation into NGN2 -iNs followed by GFP, CUX2 or ATF4 overexpression. Immunocytochemistry analysis of GFP or mCherry and 53BP1 with quantification of 53BP1 + foci. n = 5 (GFP), n = 3 (CUX2) and n = 4 (ATF4). c , Promoter luciferase reporter assays in HEK293T cells at day (D) 2 after plating, 24 h after vector transfection. luc2 ( n = 5) and RPA3 - luc2 ( n = 6). d , Experimental schematic and 53BP1 immunocytochemistry in CUX2-OE NGN2 -iNs with control or RPA3 siRNA treatment. n = 3. e , MTT viability assays after siRNA or TBHP treatment in GFP- or CUX2-OE NGN2 -iNs. n = 6 (GFP-OE and CUX2-OE + RPA3 -siRNA groups) and n = 9 (CUX2-OE + control siRNA). f , NHEJ reporter assays in GFP-, CUX2-, ATF4- or CIRBP-overexpressing hTERT RPE-1 cells or WT cells treated with siRNA (on day 0) or 1 μM AZD7648 DNA-PK inhibitor (DNA-PKi; on day 3) with quantification of NanoLuc luciferase activity normalized to the internal firefly luciferase control ( n = 4). g , Schematic of a model in which CUX2 and ATF4 proteins repair damaged DNA. Data are mean ± s.e.m. Statistical differences were determined using two-way ANOVA with Dunnett’s multiple-comparison test ( a ), one-way ANOVA with Šídák’s multiple-comparison test ( b , e and f ), Kruskal–Wallis test with Dunn’s multiple-comparison test ( c ) or two-tailed unpaired t -tests ( d ). Scale bars, 10 µm ( a , b and d ).

Article Snippet: Non-blunt NHEJ reporter plasmids were a gift from E. Rajendra (Artios Pharma) and prepared by I-SceI digestion and purification as described previously .

Techniques: Alkaline Single Cell Gel Electrophoresis, SYBR Green Assay, Staining, Over Expression, Cell Differentiation, Immunocytochemistry, Luciferase, Plasmid Preparation, Transfection, Control, Activity Assay, Comparison, Two Tailed Test

Journal: International Journal of Molecular Medicine

Article Title: miRNA-378a-5p attenuates the development of abdominal aortic aneurysm via ABLIM1-MKL1 signaling pathways

doi: 10.3892/ijmm.2026.5768

Figure Lengend Snippet: ABLIM1 is a downstream target of miR-378a-5p and involves in abdominal aortic aneurysm development. (A) Volcano plot of differently expressed genes in GSE183464 and GSE237229 database. Venn diagram showed intersection of differentially expressed genes and predicted target genes including ABLIM1 , DDX5 and SLC7A1 . (B) RT-qPCR analysis of target genes Ablim1 , Ddx5 and Slc7a1 in the TNFα-treated vascular smooth muscle cells (n=3 per group). RT-qPCR analysis of Ablim1 , Ddx5 and Slc7a1 in the aortas of Ang II-treated mice (n=3 per group). (C) ABLIM1 expression in the aortas treated-with antagomir-NC or antagomir-378a-5p was identified using by immunofluorescence staining. ABLIM1 (red), α-SMA (green) and DAPI (blue). (D) Representative images of immunofluorescence staining identified ABLIM1 expression in aortas with angomir-NC or angomir-378a-5p. ABLIM1 (red), α-SMA (green) and DAPI (blue). (E) Conservatism analysis of the binding site for miR-378a-5p and ABLIM1 in in humans, mice and rat. (F) Luciferase activity in 293T cells transfected with mimics-miR-378a-5p together with ABLIM1 -3'UTR-wild type or mutant plasmid. Data are presented as the mean ± SEM. P-values were calculated by Student's t test (for B). ** P<0.01 vs. Control or saline or mimics-NC- ABLIM1 -3'UTR-WT. ABLIM1, actin-binding LIM protein 1; miR or miRNA, microRNA; RT-qPCR, reverse transcription-quantitative PCR; α-SMA, α-smooth muscle actin; NC, negative control; UTR, untranslated region; WT, wild-type; MUT, mutated.

Article Snippet: The 3' untranslated region (3'UTR) of mouse Ablim1 , Ddx5 (Dead-box helicase 5), Slc7a1 (Cationic amino acid transporter 1) gene was amplified and cloned into the pMIR-REPORT Luciferase (OBiO Technology Company) to construct pMIR-REPORT Luciferase 3'UTR wild-type (WT) plasmids.

Techniques: Quantitative RT-PCR, Expressing, Immunofluorescence, Staining, Binding Assay, Luciferase, Activity Assay, Transfection, Mutagenesis, Plasmid Preparation, Control, Saline, Reverse Transcription, Real-time Polymerase Chain Reaction, Negative Control