mouse monoclonal 4d11  (Novus Biologicals)

Journal: PLOS One

Article Title: Targeted knockdown of DNA methyltransferase 3a (DNMT3a) unlocks dedifferentiation and neurogenic potential in mouse retinal Müller glia

doi: 10.1371/journal.pone.0337891

Figure Lengend Snippet: (A–C) Time-course analysis of mRNA expression levels of progenitor markers Ascl1, Nestin and Lin28 , normalized to Gapdh , measured by qRT-PCR. NMDA and DNMT3a KD alone transiently upregulated these markers, with a more pronounced and sustained effect observed in the DNMT3a KD NMDA group. Data represent mean ± SEM of three independent experiments; *p < 0.05 vs. CTRL at the corresponding point (two-way ANOVA with Tukey’s post hoc test). (D) Quantification of Lin28 fluorescence intensity in arbitrary units (A.U.) from immunocytochemistry images shown in (E). Significant increases in Lin28 expression were observed in NMDA, DNMT3a KD and DNMT3a KD NMDA groups compared to CTRL (*p < 0.05 vs. CTRL; #p < 0.05 vs. NMDA; one-way ANOVA with Tukey’s post hoc test). (E) Representative phase contrast and immunofluorescence images showing DAPI (nuclei, blue) and Lin28 (green) staining at 48 h post-treatment. CTRL: control, NMDA: exposure to NMDA 100 µM, MOCK: electroporation without plasmid, dCas9: transfection of empty plasmid, CRISPRi: CRISPR interference-mediated Dnmt3a knockdown, CRISPRi NMDA: Dnmt3a knockdown with simultaneous exposure to NMDA 100 µM. Phase contrast images scale bar = 100 μm, 10X; Immunofluorescence images scale bar = 20 µm, 40X.

Article Snippet: After that the samples were incubated with following primary antibodies: mouse monoclonal anti-βlll tubulin antibody (Santa Cruz sc-80005) (1:300); rabbit polyclonal anti-GS antibody (Abcam ab73593) (1:300); mouse monoclonal anti-nestin antibody (Santa Cruz sc-23927) (1:200); rabbit polyclonal anti-Lin28 (Abcam ab46020) (1:200) diluted in 1:3 blocking solution at 4°C overnight.

Techniques: Expressing, Quantitative RT-PCR, Fluorescence, Immunocytochemistry, Immunofluorescence, Staining, Control, Electroporation, Plasmid Preparation, Transfection, CRISPR, Knockdown

Journal: Cell reports

Article Title: Paraspeckle protein NONO regulates active chromatin by allosterically stimulating NSD1

doi: 10.1016/j.celrep.2025.116247

Figure Lengend Snippet: (A) Schematic illustration of annotated and putative functional domains of NSD1. NID, nuclear receptor interaction domain; PWWP, Pro-Trp-Trp-Pro domain; RBD, RNA-binding domain; PHD, plant homeodomain; AWS, associated with SET domain (also referred to as pre-SET); SET, su(var), enhancer of zeste, trithorax domain. (B) Western blot of NSD1, GAPDH, and H3K36me2 for WT or NSD1/2-dKO HEK293T cells stably rescued with WT or NSD1 mutants. (C) Top: meta-analysis profiling of genome-wide NSD1 and H3K36me2 ChIP-seq signals within −10 kb of TSS to +10 kb of TES. Bottom: representative track images of NSD1-WT or NSD1 ΔPWWP2 in HEK293T NSD1/2-dKO rescued background. TSS, transcription start site; TES, transcription end site. (D) Western blot of NSD1, GAPDH, and H3K36me2 for WT of NSD1/2-dKO HEK293T cells stably rescued with NSD1 or NSD1 PWWP2–4A . (E) Western blot of Nestin, GAPDH, and H3K36me2 for E14-mESC cells undergoing embryoid bodies (EBs) and neural progenitor cell (NPC) differentiation. (F) NPC differentiation of E14-mESCs with indicated genotypes. Top: representative images of (EBs) undergoing NPC differentiation after 3 days of retinoic acid (RA) treatment. Bottom: quantifications of differentiating and non-differentiating EBs. Scale bars, 500 μm.

Article Snippet: Nestin mouse monoclonal 10C2 , Cell Signaling Technology , Cat# 33475.

Techniques: Functional Assay, RNA Binding Assay, Western Blot, Stable Transfection, Genome Wide, ChIP-sequencing