Journal: Current biology : CB

Article Title: Expanded satellite repeats amplify a discrete CENP-A nucleosome assembly site on chromosomes that drive in female meiosis

doi: 10.1016/j.cub.2017.06.069

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: 10× T4 DNA Ligase Buffer with 10 mM ATP , New England Biolabs , B0202S.

Techniques: Immunostaining, Recombinant, DNA Labeling, Nick Translation, DNA Ligation, Purification, SYBR Green Assay, Sequencing, Clone Assay, Plasmid Preparation, Software

Journal: Nucleic Acids Research

Article Title: Construction of a highly error-prone DNA polymerase for developing organelle mutation systems

doi: 10.1093/nar/gkaa929

Figure Lengend Snippet: Mutant frequencies and DNA polymerase error rates. Error rates in columns 5A and 5B were calculated from the data in columns 3 and 4 and Taq DNA error rates shown in brackets from: 1 the supplier (New England Biolabs) and 2 McInerney et al. ( 60 ). 3 Column 5C error rates were from scoring mutations in the alpha 1 and 5 coding regions in the cI gene (this work). Columns 6D and 6E show relative error rates based on columns 5A and C respectively. nd- not determined

Article Snippet: The Taq DNA polymerase error rate in the pH 8.8 buffer provided by the supplier (New England Biolabs) was 2.85 × 10 −4 mutations per base, which is consistent with other reports ( , ).

Techniques: Mutagenesis

Journal: The EMBO Journal

Article Title: A G1‐like state allows HIV ‐1 to bypass SAMHD 1 restriction in macrophages

doi: 10.15252/embj.201696025

Figure Lengend Snippet: A, B Microglia isolated from 8‐ to 12‐week‐old CD1 mice were stained for the macrophage markers (A) F4/80 and (B) MCM2 4 h after isolation. Scale bars: 10 μm. C Graph shows the percentage of MCM2‐positive cells from four independent experiments. On average, 2 × 10 3 cells in each experiment were recorded and analysed using Hermes WiScan cell‐imaging system and ImageJ (mean ± s.e.m.).

Article Snippet: Antibodies used were as follows: anti‐cdc2 (Cell Signaling Technology, Beverly, MA, USA); anti‐CDK2 (H‐298, Santa Cruz Biotechnology); anti‐pCDK2(Thr160) (Bioss Inc., MA, USA); anti‐CDK4 (DCS156, Cell Signaling Technology); anti‐CDK6 (B‐10, Santa Cruz Biotechnology); anti‐SAMHD1 (ab67820, Abcam, UK), beta‐actin (ab6276, abcam, UK); mouse anti‐MCM2 (BM‐28, BD Biosciences, UK); and rabbit anti‐MCM2 (SP85) from Sigma; pSAMHD1 (a kind gift from M. Benkirane) and ProSci (Poway, CA, USA); anti‐Geminin (NCL‐L‐Geminin, Leica); anti‐mouse F4/80 and CD11b (kind gift from S. Yona, UCL); anti‐human CD68, CD14, CD163, CD80, CD86, CD40 (kind gift from M. Noursadeghi).

Techniques: Isolation, Staining, Imaging

Journal: The EMBO Journal

Article Title: A G1‐like state allows HIV ‐1 to bypass SAMHD 1 restriction in macrophages

doi: 10.15252/embj.201696025

Figure Lengend Snippet: A–D Peritoneal macrophages from 6‐ to 10‐week‐old C57BL/6 mice were stained for the macrophage markers (A) F4/80, (B) CD11b, (C) MCM2 or (D) active DNA synthesis (EdU) 2 h after isolation. Scale bars: 20 μm. E–K Peritoneal macrophages were isolated from 6‐ to 10‐week‐old WT or SAMHD1 −/− (KO) mice. Cells were infected with VSV‐G‐pseudotyped HIV‐1 GFP for 48 h and analysed for (G) infection, (H) MCM2 expression, (I) EdU incorporation (EdU was added at the time of infection), and co‐localisation between infection and (J) MCM2 protein or (K) active DNA synthesis (by EdU incorporation). (E, F) Random microscopic fields and plot profiles show an example of immunofluorescence intensity along infected cells. Scale bars: 10 μm. On average, 10 4 cells in each experiment were recorded and analysed using Hermes WiScan cell‐imaging system and ImageJ ( n ≥ 2, mean ± s.e.m.; (ns) non‐significant; **P ‐value ≤ 0.01, unpaired t ‐test).

Article Snippet: Antibodies used were as follows: anti‐cdc2 (Cell Signaling Technology, Beverly, MA, USA); anti‐CDK2 (H‐298, Santa Cruz Biotechnology); anti‐pCDK2(Thr160) (Bioss Inc., MA, USA); anti‐CDK4 (DCS156, Cell Signaling Technology); anti‐CDK6 (B‐10, Santa Cruz Biotechnology); anti‐SAMHD1 (ab67820, Abcam, UK), beta‐actin (ab6276, abcam, UK); mouse anti‐MCM2 (BM‐28, BD Biosciences, UK); and rabbit anti‐MCM2 (SP85) from Sigma; pSAMHD1 (a kind gift from M. Benkirane) and ProSci (Poway, CA, USA); anti‐Geminin (NCL‐L‐Geminin, Leica); anti‐mouse F4/80 and CD11b (kind gift from S. Yona, UCL); anti‐human CD68, CD14, CD163, CD80, CD86, CD40 (kind gift from M. Noursadeghi).

Techniques: Staining, DNA Synthesis, Isolation, Infection, Expressing, Immunofluorescence, Imaging