Journal: The Journal of Neuroscience

Article Title: Cenpj Regulates Cilia Disassembly and Neurogenesis in the Developing Mouse Cortex

doi: 10.1523/JNEUROSCI.1849-18.2018

Figure Lengend Snippet: Cenpj was specifically depleted in the dorsal telencephalon of CenpjCKO mice. A, Schematic of Cenpj-trapped mice construction strategy. A LacZ reporter cassette and a neo tag, flanked by two FRT sequences, were introduced between the fourth exon and the fifth exon of Cenpj gene locus. The fifth exons were flanked by two LoxP sites in CenpjLacZ mice. LacZ reporter cassettes can be removed by Flippase splicing and the Cenpj gene can be knocked out by tissue-specific Cre recombinase splicing. B, Western blot analysis for Cenpj +/+ and CenpjCKO cerebral cortex lysate at E15.5. C, Quantification of Cenpj knock-out by Western blot. Histogram shows the mean ± SD; ****p < 0.0001 as determined by a t test; n = 3. D, Brain sections were immunostained by anti-Pericentrin (green) antibody and anti-Cenpj (red) antibody on the ventricular surface in Cenpj+/+ and CenpjCKO mice. Scale bar, 10 μm. E, Representative whole-mount images of Cenpj+/+ and CenpjCKO brains at E15.5. Scale bar, 0.5 mm. F, Quantification of cerebral hemisphere circumference at E15.5. Data are presented as the mean ± SD; ***p = 0.0004 as determined by a t test; n = 3. G, Representative images of Cenpj+/+ and CenpjCKO brain coronal sections at E15.5 by Nissl staining. Right, Magnified cortical column. Scale bars, 500 μm (left) and 100 μm (right). H, Histogram of the cerebral cortex thickness. Data are presented as the mean ± SD; ****p < 0.0001 as determined by a t test; n = 3; 3 brain slices per experiment. I, Body size of Cenpj+/+ and CenpjCKO mice at P7. J, Representative images of the Cenpj+/+ and CenpjCKO brain size at P7. Scale bar, 1 mm. K, Quantification of P7 cerebral hemisphere circumference. Data are presented as the mean ± SD; ****p < 0.0001 as determined by a t test; n = 3. L, Representative images of Ctip2 (green) and Satb2 (red) expressions in the Cenpj+/+ and CenpjCKO cortex at P7. High-magnification images show the cortical column. Scale bar, 30 μm. M, Quantification of Ctip2+ cells and Satb2+ cells ratios in the neocortex. Data are presented as the mean ± SD; *pCtip2 = 0.0335, **pSatb2 = 0.0011 as determined by a t test; n = 3; 3 brain slices per experiment. N, Representative images of Foxp2 and Cux1 expressions in the Cenpj+/+ and CenpjCKO cortex at P7. Scale bar, 30 μm. O, Quantification of Foxp2+ cells ratios in the neocortex. Data are presented as the mean ± SD; *p = 0.0175, as determined by a t test; n = 3; 3 brain slices per experiment. P, Quantification of Cux1+ cells ratios in the neocortex. Data are presented as the mean ± SD; ***p = 0.0001 as determined by a t test; n = 3; 3 brain slices per experiment.

Article Snippet: Primary antibodies used were as follows: Zo-1 (1:300, 339100; Invitrogen), γ-Tubulin (1:5000, T3559; Sigma-Aldrich), γ-Tubulin (1:5000, T5326; Sigma-Aldrich), Acetylated Tubulin (1:30000, T7451; Sigma-Aldrich), Cenpj (1:50, 11517-1-AP; Proteintech), Pericentrin (1:300, 611814; BD Biosciences), GFP (1:500, GFP-1020; Aves Laboratories), Cleaved Caspase 3 (1:300, 9664; Cell Signaling Technology), Tbr2 (1:300, ab23345; Abcam), Satb2 (1:300, ab34735; Abcam), Ctip2 (1:300, ab18465; Abcam), Ki67 (1:300, ab9260; Millipore), BrdU (1:300, ab6326; Abcam), Pax6 (1:300, PRB-278P; Covance), Sox2 (1:300, sc-17320; Santa Cruz Biotechnology), EdU (Click-iT EdU Alexa Fluor 594 Imaging Kit, {"type":"entrez-nucleotide","attrs":{"text":"C10339","term_id":"1535410","term_text":"C10339"}} C10339 ; Thermo Scientific), GFAP (1:500, G9269,Sigma-Aldrich), and β-Catenin (1:300, 610153; BD Biosciences).

Techniques: Western Blot, Knock-Out, Staining

Journal: Cell reports

Article Title: BRCA1 Mutation-Specific Responses to 53BP1 Loss-Induced Homologous Recombination and PARP Inhibitor Resistance

doi: 10.1016/j.celrep.2018.08.086

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit Anti-CtIP Polyclonal antibody , Bethyl , Cat# A300–488A; RRID: AB_2175262.

Techniques: Labeling, Polymer, Recombinant, Plasmid Preparation, Staining, Mutagenesis, Extraction, Expressing, TaqMan Assay, shRNA, Software

Journal: Nature communications

Article Title: PARP3 is a promoter of chromosomal rearrangements and limits G4 DNA.

doi: 10.1038/ncomms15110

Figure Lengend Snippet: Figure 7 | PARP3 and BLM cooperate to repair DNA DSBs. (a–c) Quantification of IF for 53BP1 (a) and gH2AX (b) at the indicated time points in wild- type (WT) or PARP3 / A549 cells transfected with siRNA targeting control (siCont) or BLM (siBLM). Data are represented as mean±s.e. of three biological replicates. P values calculated using unpaired Student’s t-test. *Po0.05, **Po0.01, ***Po0.001. (c) immunoblot after transfection with siRNA targeting control (Ct) or BLM.

Article Snippet: Equivalent amounts of chromatin were incubated with primary antibody (overnight at 4 C for RPA, CtIP, and BLM and HA, 2 h at 4 C for gH2AX) followed by protein-G magnetic beads (Cell Signaling Technology).

Techniques: Transfection, Control, Western Blot