Journal: PLoS ONE

Article Title: Characterization of the Molecular Mechanism of the Bone-Anabolic Activity of Carfilzomib in Multiple Myeloma

doi: 10.1371/journal.pone.0074191

Figure Lengend Snippet: ( A – D ) Cells were treated with 20-nM CFZ for indicated times, and nuclear fractions were extracted from cell lysates and subjected to GST-E-cadherin pull-down assays. AB818 protein was measured as a loading control. ( A ) Proteins from indicated OB and MSC cell lines were immunoblotted with anti-β-catenin antibody or ( B ) with an antibody that specifically recognizes active, non-phosphorylated β-catenin. ( C ) Protein from human MSCs from a healthy donor or ( D ) from bone marrow of a patient with MM were immunoblotted with anti-β-catenin antibody (F-β-cat) or with an antibody that specifically recognizes active, non-phosphorylated β-catenin (A-β-cat). ( E ) HS5 cells, ( F ) indicated cell lines, or ( G ) human MSCs from a healthy donor (NMSC) or from bone marrow of a patient with MM (MMMSC) were treated with indicated concentrations of CFZ for 12 hours, and β-catenin in nuclei and cytoplasm of the cells was determined with analysis of immunofluorescence staining, using an antibody specific for active β-catenin. Images were taken with a fluorescence microscope, as described in Methods.

Article Snippet: Fluorescence was detected with an Axio Observer A1 fluorescence microscope with 10X objective lens (Carl Zeiss Microscopy, Jena, Germany) and images were captured with a SPOT camera (Diagnostic Instruments, Sterling Heights, MI).

Techniques: Control, Immunofluorescence, Staining, Fluorescence, Microscopy

Journal: Scientific Reports

Article Title: The Role of AMPARs in the Maturation and Integration of Caudal Ganglionic Eminence-Derived Interneurons into Developing Hippocampal Microcircuits

doi: 10.1038/s41598-019-41920-9

Figure Lengend Snippet: The density of dendritic but not somatic glutamatergic synapses is determined by the presence or absence of GluA2. ( A , B ) Images show (from two different angles) Airyscan (LSM) super-resolution images of green fluorescent puncta (PSD-95_EGFP) detected on magenta fluorescent (tdTomato) dendrites and somata converted using Imaris software (Bitplane). PSD-95_EGFP expression that created a punctate signal on the dendrites and soma, is used as a proxy for glutamatergic synaptic sites in 3D space. The tdTomato signal was used to create a surface for dendrites and soma and the EGFP signal that did not fall within 1 µm distance from the created surface was first automatically and then, if necessary, manually eliminated from the analysis. Scale bars, 3 µm. small panels to the right represent the raw images used for analysis. ( C ) Bar graphs of the density of synaptic sites on dendrites (WT, n = 64; GluA2 KO, n = 30; GluA1-3 KO, n = 34) or soma (WT, n = 27; GluA2 KO, n = 22; GluA1-3 KO, n = 13) of the CGE interneurons. The data for all three groups were compared for significance with ANOVA and a posthoc Wilcoxon-Mann-Whitney Test when a significance was found with ANOVA. The density of synaptic sites on soma are similar in all groups compared. The density of synaptic sites is significantly reduced on KO CGE interneurons compared to WT: GluA2 KO to WT, p = 0.034 × 10 −8 , GluA1-3 KO to WT, p = 0.024 × 10 −8 , GluA1-3 to GluA2, p = 0.53. ***p < 0.0005.

Article Snippet: Fluorescent images were captured using the 10X objective of a Zeiss LSM fluorescent microscope and 10 images of hippocampi from each mouse were used to count.

Techniques: Software, Expressing, MANN-WHITNEY