Journal: Cell reports

Article Title: miRNA551b-3p Activates an Oncostatin Signaling Module for the Progression of Triple-Negative Breast Cancer

doi: 10.1016/j.celrep.2019.11.085

Figure Lengend Snippet:

Article Snippet: T47D , ATCC , Cat# 85102201; RRID: CVCL_0553.

Techniques: Purification, Recombinant, Transfection, Magnetic Beads, Protease Inhibitor, Modification, Bicinchoninic Acid Protein Assay, Fractionation, Cell Culture, Enzyme-linked Immunosorbent Assay, SYBR Green Assay, Reporter Assay, Immunoprecipitation, Plasmid Preparation, Northern Blot, Negative Control, Mutagenesis, Expressing, Software, CRISPR

Journal: Nature neuroscience

Article Title: Social Deficits in Shank3 -deficient Mouse Models of Autism Are Rescued by histone deacetylase (HDAC) Inhibition

doi: 10.1038/s41593-018-0110-8

Figure Lengend Snippet: ( a ) Immunoblots and quantification analysis of the level of acetylated H3 and total H3 in the nuclear fraction of cortical slices from wild-type (WT) or Shank3 +/ΔC (Het) mice injected (i.p.) with saline or romidepsin (RMD, 0.25 mg/kg, 3×). Immunoblotting was performed at 4-5 days post-injection. F 1,20 =11.3, P =0.0031; * P <0.05, two-way ANOVA, n=6 each group. ( b, c ) Plots showing the time spent investigating either the social (Soc) or nonsocial (NS) stimulus (b) and the social preference index (c) during 3-chamber sociability testing of saline-injected WT (n=18), saline-injected Shank3 +/ΔC (n=17), RMD-treated Shank3 +/ΔC (n=17) and RMD-treated WT (n=12) mice. In (b), F 3,120 =11.4, P <0.0001; +++ P <0.001 (Soc vs. NS), ** P <0.01, *** P <0.001, two-way ANOVA. In (c), F 1,60 =58.5, P <0.0001; *** P <0.001, two-way ANOVA. ( d ) Representative heat maps illustrating the time spent in different locations of the 3 chambers from the social preference tests of all groups. Locations of Sol and NS stimuli are labeled with the circles. ( e ) Plots of social preference index in Shank3 +/ΔC mice treated with romidepsin (n=10) or saline (n=10) at different time points. F 1,18(treatment) =124.3, P <0.0001; *** P <0.001 (saline vs. romidepsin), ### P <0.001 (pre- vs. post-injection), two-way rmANOVA. ( f ) Plots of social preference index in Shank3 +/ΔC mice treated with different doses of romidepsin (0.025 mg/kg, n=9; 1 mg/kg, n=8) at different time points. F 1,15(treatment) =51.3, P <0.0001; *** P <0.001 (1 mg/kg vs. 0.025 mg/kg RMD); ### P <0.001 (pre- vs. post-injection), two-way rmANOVA. ( g ) Scatter plots showing total sniffing time in social approach tests of saline-injected WT (n=12), saline-injected Shank3 +/ΔC (n=14), RMD-treated Shank3 +/ΔC (n=12) and RMD-treated WT (n=12) mice. F 1,46 =6.0, P =0.018; * P <0.05, ** P <0.01, two-way ANOVA. ( h ) Representative heat maps illustrating the time spent in different locations of the apparatus from the social approach tests of all groups. Locations of social stimuli are labeled with the circles. ( i-n ) Plots of social preference index in Shank3 +/ΔC mice treated with fluoxetine (10 mg/kg, i.p., 14×, i, n=9), clozapine (5 mg/kg, i.p., 3×, j, n=11), valproic acid (VPA, 100 mg/kg, i.p., 3×, k, n=11), aripiprazole (1 mg/kg, i.p., 3×, l, n=9), risperidone (0.1 mg/kg, i.p., 3×, m, n=10), or Trichostatin A (TSA, 0.5 mg/kg, i.p., 3×, n, n=8). F 2,30 =19.2 (VPA), P <0.0001; F 2,21 =19.7, P <0.0001 (TSA); ### P <0.001 (pre- vs. post-injection), one-way ANOVA. ( o ) Representative heat maps illustrating the time spent in different locations of the 3 chambers from the social preference tests of a Shank3 e4-9 mouse before and after romidepsin treatment (0.25 mg/kg, i.p., 3×). ( p ) Box plots showing the time spent investigating either Soc or NS stimulus during sociability testing in WT (n=8) or homozygous Shank3 e4-9 mice (n=10) before and after romidepsin treatment. F 2,50 =9.2, P =0.0003; +++ P <0.001 (Soc vs. NS), * P <0.05, *** P <0.001, two-way ANOVA. ( q ) Scatter plots showing the preference index of the sociability testing in individual Shank3 e4-9 mice before and after romidepsin treatment (n=10). t 9 =4.36, ** P =0.0018, paired two-tailed t -test. ( r ) Plots of social preference index in Shank3 e4-9 mice (n=10) treated with romidepsin at different time points. F 6,63 =11.8, P <0.0001; * P <0.05, ** P <0.01, *** P <0.001 (pre- vs. post-injection), one-way ANOVA. Shank3 +/ΔC mice (a-n) and WT mice (a-n,p) are all males (5-6 weeks old); Shank3 e4-9 mice (o-r) are 6 males and 4 females (5-6 weeks old). Data are presented as median with interquartile range (a,b,p) or mean ± SEM (c,e-g,i-n,q,r). Each set of the experiments was replicated for at least 3 times. See for blot source data.

Article Snippet: Romidepsin (Selleckchem), Fluoxetine (NIH), Valproic acid (Tocris), aripiprazole (Tocris), risperidone (Tocris), or Trichostatin A (Sigma) was prepared by dissolving in DMSO to make the stock solution and diluting with 0.9% saline before injections (DMSO concentration of the working solution: <0.2%).

Techniques: Western Blot, Injection, Labeling, Two Tailed Test