Journal: Journal of Biological Chemistry

Article Title: The Insulin Receptor Substrate of 53 kDa (IRSp53) Limits Hippocampal Synaptic Plasticity

doi: 10.1074/jbc.m808425200

Figure Lengend Snippet: FIGURE 7. Behavioral analysis. A and B, open field test. Animals (n 10 per genotype) were placed into the open field box, and the total distance moved (A) as well as the time spent in the center of the box (B) were recorded for three subsequent 5-min intervals. C, Rotarod test. Animals were placed on an accelerated rotating corrugated rod; during four trials (trial 1–3 on the first day; trial 4 on the next day) lasting 300 s each, the latency of mice until they fall from the rod was recorded. D, contextual fear-conditioning test. Mice were placed into a boxwithametalgridfloor(context);electro-shockswereapplied,whereasthetimethatmicespentmotionless (freezing) was continuously monitored during the 2 min before (Base line) and 2 min after the first foot shock (Aftershock).Onthenextdaythemicewerereturnedtothesameboxwithoutapplyinganyshock,andthetime spent freezing was again monitored for 3 min (Retrieval). Note the significant difference between WT (/) and KO (/) mice during retrieval, indicating that KO mice do not learn or memorize the association between the foot shocks and the context (***, p 0.0001, Newman-Keuls post-hoc after two-way analysis of variance for repeated measurements).

Article Snippet: In the accelerated Rotarod testmice had towalk on a turning, corrugated rod (3.2 cm in diameter) (Acceler Rotarod for mice Jones & Roberts, TSE Systems, Bad Homburg, Germany).

Techniques:

Journal: Oncotarget

Article Title: Melanoma-initiating cells exploit M2 macrophage TGFβ and arginase pathway for survival and proliferation

doi:

Figure Lengend Snippet: Data for all bar graphs are expressed as fold change in sphere-forming efficiency relative to untreated CD34 − TICs. Bars represent mean ± SE, * P < 0.05, **P < 0.01, ***P < 0.001, **** P < 0.0001, N.S. not significant, 1-way ANOVA for B and D to F, 2-way ANOVA for C. (A) Flow cytometry sorting of immune cell populations. The Lin + gate contain T, B and NK cells. Lin − cells are separated into CD11b + CD68 − myeloid cells and CD11b + CD68 + TAMs. (B) Effect of immune cell populations on sphere formation: CD34 − TICs were cultured with the immune cell populations sorted in A. (C) Effect of TAM inhibition on sphere formation: CD34 − TICs were cultured with or without TAMs. Two CSF1R inhibitors Ki20227 (Ki; 100nM) and CD115-neutralizing antibody (α-CD115; 1μg/ml) were added for the duration of the culture. (D) Effect of BMDM on sphere formation: BMDM from tumor-bearing mice (RET + ) and non-tumor bearing littermates (ret − ) were polarized to M1 or M2 phenotype and co-cultured with CD34 − TICs. (E) Effect of conditioned medium (CM) on sphere formation: CM generated from M1- or M2-polarized BMDM were added to CD34 − TICs at 1/10 th of stem cell media volume. (F) Effect of CSF1R inhibitors on the stimulatory effect of M2-BMDM CM (M2-CM): Ki20227 (100nM) and anti-CD115 (1μg/ml) were either added to the BMDM prior to CM generation (Ki20227 > CM, α-CD115 > CM) or added to the CM during sphere culture (CM+Ki20227, CM+α-CD115).

Article Snippet: Mice underwent gavage with Ki20227 (30mg/kg/day in 0.05% methylcellulose; Accelachembio) over 10 days.

Techniques: Flow Cytometry, Cell Culture, Inhibition, Generated

Journal: Oncotarget

Article Title: Melanoma-initiating cells exploit M2 macrophage TGFβ and arginase pathway for survival and proliferation

doi:

Figure Lengend Snippet: Data for D to F are expressed as fold change in sphere-forming efficiency relative to untreated CD34 − TICs. Statistical analysis: **P < 0.01, **** P < 0.0001, N.S. not significant. (A) Principle Component Analysis (PCA) mapped scatter plot: The global gene expression profiles of untreated (red) or M2-CM treated CD34 − spheres (green) and CD34 + spheres (black) were analyzed by PCA. Graph represents the first three principal components of the microarray analysis data (PC1, PC2 and PC3) in the X, Y and Z axis respectively. (B) Comparison of number of tumor nodules formed in lungs of Rag1 mice injected with CD34 − spheres derived from untreated, M2-CM- or TAM-stimulated cultures. Twenty lung sections were sampled from each mouse. Each point represent one mouse, Wilcoxon matched-pairs signed rank test. (C) Comparison of size of the tumor nodules presented in B. Bars represent geometric mean ± 95% CI, 1-way ANOVA. (D) Effect of in vivo CD115 inhibition on sphere formation in vitro : CD34 − TICs from mice treated with vehicle, Ki20227 or anti-CD115 for 10 days were cultured with or without TAMs from untreated mice. Bars represent mean ± SE, 2-way ANOVA. (E) Effect of in vivo CD115 inhibition on TAM activity in vitro : CD34 − TICs from untreated mice were cultured with TAMs from untreated (+UT) mice, or mice treated with vehicle (+Veh), Ki20227 for 10 days (+Ki) or Ki20227 for 10 days followed by 10 days recovery [+Ki (re)] mice. Bars represent mean ± SE, 1-way ANOVA. (F) Recovery of macrophage-responsive TICs: CD34 − TICs from vehicle-, Ki20227- or Ki20227 recovery- treated mice were cultured with or without TAMs from untreated mice. Bars represent mean ± SE, 2-way ANOVA.

Article Snippet: Mice underwent gavage with Ki20227 (30mg/kg/day in 0.05% methylcellulose; Accelachembio) over 10 days.

Techniques: Gene Expression, Microarray, Comparison, Injection, Derivative Assay, In Vivo, Inhibition, In Vitro, Cell Culture, Activity Assay