Journal: Scientific Reports

Article Title: A novelty-retrieval-extinction paradigm leads to persistent attenuation of remote fear memories

doi: 10.1038/s41598-020-60176-2

Figure Lengend Snippet: Exposure to an OF before the retrieval-extinction session increased hippocampal H3K9/14 acetylation. ( A ) Representative images showing immunostaining for H3K9/14ac in the hippocampal CA3 region 1 h after completion of extinction in nov-Ret and control mice. Higher magnification images of H3K9/14ac staining in the CA3 are shown for the control (b) and nov-Ret groups (d) and correspond to the labelled regions in a and c, respectively. Scale bar: 100 μm. ( B ) Immunohistochemical analysis of OD for H3K9/14ac in the hippocampus for each group. n = 4 mice in each group. ( C , E ) Pictures of western blotting analysis of H3K9/14ac in the hippocampus 1 h after completion of extinction in control and nov-Ret mice. control, n = 5; Ret, n = 2; nov, n = 2; nov-Ret, n = 6. Two mice in the nov-Ret group received fear conditioning and retrieval-extinction in the same context (AA), and another four in different contexts (AB). Original western blotting images are shown in Supplementary Fig. . ( D , F ) Densitometric analysis for H3K9/14ac in ( C , E ), respectively. Data are presented as mean ± SEM. ** P < 0.01, *** P < 0.001 vs . control. OF, open field; SEM, standard error of mean.

Article Snippet: The membranes were blocked with 5% non-fat milk for 2 h at room temperature and incubated overnight with the following antibodies: rabbit polyclonal anti-histone H3 antibody (Servicebio; dilution, 1:30000) and rabbit polyclonal antibody H3K9/14ac (Origene; dilution, 1:1000).

Techniques: Immunostaining, Staining, Immunohistochemical staining, Western Blot

Journal: Cell Discovery

Article Title: A genome-scale CRISPR-Cas9 screening method for protein stability reveals novel regulators of Cdc25A

doi: 10.1038/celldisc.2016.14

Figure Lengend Snippet: Acetylation of lysine 150 on Cdc25A negatively regulates the G2/M checkpoint in response to IR. ( a ) Knockout of endogenous Cdc25A in HeLa cells through CRISPR-Cas9. ( b ) Re-expression of wild-type (WT) or K150R mutant Cdc25A in the Cdc25A knockout cell line through pSIN lentivirus. ( c, d ) Degradation rate of WT-Cdc25A and K150R-Cdc25A mutant post IR (6 Gy) using western blot. ( e, f ) Cdc25A KO HeLa cells stably re-expressing WT-Cdc25A or K150R-Cdc25A mutant were exposed to IR (6 Gy). The cells were harvested and analyzed using flow cytometry after incubation with 100 ng ml −1 of nocodazole for 10 h. The percentage of cells positive for phospho-histone H3 (p-H3) is indicated. N =3. Bars indicate the s.e.m. *** P <0.001, Student’s t- test.

Article Snippet: After the cells were centrifuged, the cell pellet was suspended in 100 ml of PBS containing 1% bovine serum albumin and 0.75 μg of a polyclonal antibody that specifically recognizes the phosphorylated form of histone H3 (Upstate Biotechnology, Lake Placid, NY, USA), and incubated for 1.5 h at room temperature.

Techniques: Knock-Out, CRISPR, Expressing, Mutagenesis, Western Blot, Stable Transfection, Flow Cytometry, Incubation