Journal: The FEBS journal

Article Title: A fluorinated indole-based MDM2 antagonist selectively inhibits the growth of p53 wt osteosarcoma cells

doi: 10.1111/febs.14774

Figure Lengend Snippet: The summary of activities of the trifluoro-substituted compounds. FP – fluorescence polarization assay, NMR - 1 H- 15 N HSQC NMR, MST - microscale thermophoresis.

Article Snippet: Human osteosarcoma SJSA-1 (p53 wt ) cell line was purchased from ATCC (LGC Standards) and cultured in RPMI-1640 Medium containing L-glutamine and supplemented with 10% FBS.

Techniques: FP Assay, Nuclear Magnetic Resonance, Microscale Thermophoresis, Activity Assay

Journal: The FEBS journal

Article Title: A fluorinated indole-based MDM2 antagonist selectively inhibits the growth of p53 wt osteosarcoma cells

doi: 10.1111/febs.14774

Figure Lengend Snippet: (A,B) Stereoview of (R)-6a in complex with MDM2 (residues 18–125) – cartoon and surface representation. Inhibitor shown as green sticks, MDM2 shown in grey, waters showed as red spheres; (C) Comparison to p53 peptide, PDB ID 1YCR (shown as magenta ribbon, key residues shown as sticks).

Article Snippet: Human osteosarcoma SJSA-1 (p53 wt ) cell line was purchased from ATCC (LGC Standards) and cultured in RPMI-1640 Medium containing L-glutamine and supplemented with 10% FBS.

Techniques: Comparison

Journal: The FEBS journal

Article Title: A fluorinated indole-based MDM2 antagonist selectively inhibits the growth of p53 wt osteosarcoma cells

doi: 10.1111/febs.14774

Figure Lengend Snippet: (A,B) Western blot analysis of the expression of p53, p21 and Mdm2 in U-2 OS cells following the treatment with the indicated compounds. DMSO-treated cells served as a control (marked with “-”). (C) The expression of p53-target genes (MDM2 and CDKN1A) and the gene encoding p53 protein (TP53). U-2 OS cells were treated with 10 μM of the compound (R)-5a or 5 μM RG7388 for 6 hours. DMSO was used as a control. The expression of genes was quantified by real-time PCR with the use of Taq-Man probes. The results show mean+SEM values from three experiments, each normalized to GAPDH expression. The statistical significance was evaluated using a t-test: * p<0.05, ** p<0.01, *** p<0.001. (D) To test cell viability, MTT assay was performed. U-2 OS (p53wt), SJSA-1 (p53wt) and SAOS-2 (p53del) cells were treated with the indicated concentrations of the tested compounds and cultured for 5 days. The graph shows cell viability normalized to DMSO-treated control cell. (E) Cell cycle analysis of U-2 OS cells treated with the indicated compounds or with DMSO (control). The cells were treated for 24 hours with BrdU pulse-labeling for the last 1 hour of the treatment. The cells were stained with FITC-conjugated anti-BrdU antibody and propidium iodide (PI). The percentages of the cells in the S-phase are given on the plots. (F) Caspase 3/7 activity assay. SJSA-1 and U-2 OS cells were treated with the indicated concentrations of RG7388, (R)-5a, (R)-6a or DMSO for 24 or 48 hours. Staurosporine, a potent inducer of cell apoptosis, was used as a positive control. The results show mean+SEM values from three experiments, each normalized to DMSO-treated control cells. The statistical significance was evaluated using a t-test versus DMSO-treated control cells: * p<0.05, ** p<0.01, *** p<0.001.

Article Snippet: Human osteosarcoma SJSA-1 (p53 wt ) cell line was purchased from ATCC (LGC Standards) and cultured in RPMI-1640 Medium containing L-glutamine and supplemented with 10% FBS.

Techniques: Western Blot, Expressing, Control, Real-time Polymerase Chain Reaction, MTT Assay, Cell Culture, Cell Cycle Assay, Labeling, Staining, Activity Assay, Positive Control

Journal: NeuroImage

Article Title: Ex Vivo Fetal Brain MRI: Recent Advances, Challenges, and Future Directions

doi: 10.1016/j.neuroimage.2019.03.034

Figure Lengend Snippet: Note that regions with FA values lower than 0.15–0.2 are observed in the majority (40 GW) and peripheral regions (3 years old) of the arcuate fasciculus that cannot be detected with popular DTI analysis methods with an FA threshold (Modified from Wilkinson et al., 2017, with permission). In addition to other technical issues including the quality of MRI scan acquisition and analysis, thresholding parameters based on FA values are important factors when performing tractography.

Article Snippet: The structural MRI scan is made available by the Allen Institute for Brain Science ( Miller et al. , 2014 ), and is available at: http://download.alleninstitute.org/brainspan/MRI_DTI_data_for_prenatal_specimens/ .

Techniques: Modification

Journal: NeuroImage

Article Title: Ex Vivo Fetal Brain MRI: Recent Advances, Challenges, and Future Directions

doi: 10.1016/j.neuroimage.2019.03.034

Figure Lengend Snippet: The GE wanes between GW 20 and 30. Coronal planes from a structural T1w MRI scans of a human at GW19 show cell dense regions consisting of the GE and cortical proliferative pool (CPP). Comparing the growth of these two proliferative pools throughout development in humans, macaques, and mice permits identifying evolutionary changes in neurogenesis timing across species. The structural MRI scan is made available by the Allen Institute for Brain Science (Miller et al., 2014), and is available at: http://download.alleninstitute.org/brainspan/MRI_DTI_data_for_prenatal_specimens/. Image credit: Allen Institute.

Article Snippet: The structural MRI scan is made available by the Allen Institute for Brain Science ( Miller et al. , 2014 ), and is available at: http://download.alleninstitute.org/brainspan/MRI_DTI_data_for_prenatal_specimens/ .

Techniques:

Journal: NeuroImage

Article Title: Ex Vivo Fetal Brain MRI: Recent Advances, Challenges, and Future Directions

doi: 10.1016/j.neuroimage.2019.03.034

Figure Lengend Snippet: (A) The timing of developmental transformations (expressed as age in days after conception in macaques and humans) is regressed against equivalent developmental transformations found in mice (Workman et al., 2013; Charvet et al., 2017b, Charvet and Finlay, 2018; Charvet et al., 2018). The timing of developmental transformations is instrumental in identifying corresponding ages across species. Such an approach permits identifying variation in the timing of select developmental processes after controlling for variation in developmental schedules across species. Using this approach, Charvet et al. (2017a) found that the ganglionic eminence (GE) (B) and cortical proliferative pool (C) grow for significantly longer in humans and macaques once overall differences in the duration of developmental schedules are controlled for (see Workman et al., 2013; Charvet et al., 2017a; Charvet and Finlay, 2018; Charvet et al., 2018). (D) Examples of equivalent developmental time points are shown from reconstructed structural MRI scans and micro CT-scans. Micro CT-scans of prenatal mice are from Wong et al., 2012. Structural MR scans of prenatal humans are from the multi-dimensional human embryo project (http://embryo.soad.umich.edu/index.html).

Article Snippet: The structural MRI scan is made available by the Allen Institute for Brain Science ( Miller et al. , 2014 ), and is available at: http://download.alleninstitute.org/brainspan/MRI_DTI_data_for_prenatal_specimens/ .

Techniques: Micro-CT