Journal: Human Reproduction (Oxford, England)

Article Title: Subphenotype meta-analysis of testicular cancer genome-wide association study data suggests a role for RBFOX family genes in cryptorchidism susceptibility

doi: 10.1093/humrep/dey066

Figure Lengend Snippet: Genes containing suggestive intragenic markers common to undescended testis (UDT) and testicular cancer consortium (TECAC) case–case and case–control GWAS analyses.

Article Snippet: We quantified transcript expression by quantitative reverse transcriptase-PCR (qRT-PCR) using a protocol reported previously ( Johnson et al. , 2010 ; Barthold et al. , 2008 , 2013 ) and TaqMan gene expression assays (Thermo Fisher Scientific, USA) specific for Rbfox1 (Rn01761235_m1), Rbfox2 (Rn01197021_m1), Rbfox3 (Rn01464214_m1) and Gapdh (Rn99999916_s1).

Techniques: Expressing, Protein Binding, Migration, Activation Assay, Sterility, Modification, Transduction

Journal: MRS bulletin

Article Title: Functional imaging of brain organoids using high-density microelectrode arrays.

doi: 10.1557/s43577-022-00282-w

Figure Lengend Snippet: Figure 2. Human cerebral organoids comprise diverse cell types relevant to the study of neuronal circuit development and physiology. (a) Immunohistochemistry (IHC) revealed the presence of mature neurons (Tau/NeuN), astrocytes (GFAP), and oligodendrocytes (OLIG2) after 100 days of differentiation; nuclei were stained with DAPI (in blue). Scale bars are 50 μm. (b) Quantification of NeuN-positive cells. The ratio of the number of NeuN-positive cells to the total number of cells was quantified for both hCO lines (3 hCOs per line, for each hCO > 5000 cells). The ratio of NeuN-positive cells to total cells did not differ significantly between the two lines. (c) Single-cell RNA sequencing (scRNA- seq) was performed to quantify the cellular composition of hCOs and their comparability. The results are summarized in two pie charts with the identified cell type fractions displayed for each line. For both lines the results confirmed the presence of dopaminergic, GABAergic, glutamatergic and cholinergic neurons, neural crest cells (NCCs), radial glia (RG), astrocytes and oligodendrocytes.

Article Snippet: The following primary and secondary antibodies were used in this study: Pax6 (BioLegend, San Diego, CA, USA, #901301, 1:300), Sox2 (Sigma-Aldrich, AB5603, 1:300), FoxG1 (Abcam, ab18259, 1:200), Tuj1 (BioLegend, #801202, 1:800), Ctip2 (Abcam, ab18465, 1:200), Tbr1 (Abcam, ab31940, 1:300), MAP2 (Thermo Scientific, PA1-10,005, 1:800), Tau (Thermo Scientific, MN1000, 1:500), NeuN (Boster Bio, Pleasanton, CA, USA, M11954-3, 1:300), GFAP (Novus Biologicals, Englewood, CO, USA, NB300-141 , 1:500), goat anti-mouse IgG (H + L) highly cross-adsorbed secondary antibody, Alexa Fluor Plus 488 (Thermo Scientific, A32723, 1:400), goat anti-rabbit IgG (H + L) highly cross-adsorbed secondary Antibody, Alexa Fluor 568 (Thermo Scientific, A11036, 1:400), goat anti-rat IgG (H + L) cross-adsorbed secondary antibody, Alexa Fluor 647 (Thermo Scientific, A21247, 1:400) and goat anti-chicken IgY (H + L) cross-adsorbed secondary antibody, Alexa Fluor Plus 647 (Thermo Scientific, A32933, 1:400).

Techniques: Immunohistochemistry, Staining, RNA Sequencing

Journal: Stem Cells

Article Title: NK-cell cytotoxicity toward pluripotent stem cells and their neural progeny: impacts of activating and inhibitory receptors and KIR/HLA mismatch

doi: 10.1093/stmcls/sxae083

Figure Lengend Snippet: B7H6 expression is maintained in motoneurons differentiated from iPS-derived neural progenitor cells. (A) B7H6 (NCR3LG1) transcription was assessed by qPCR analysis using cDNA from the indicated cells, displaying relative expression using GNB2LI as housekeeping control. The tumor cell lines HCT15 and MDA.MB.231 have been included as B7H6+ and B7H6− controls, respectively. The results are normalized and represent 3 or more independent triplicate experiments. Neurons were generated from iPS-derived NPCs, and their phenotype was documented by (B) phase contrast microscopy demonstrating dendritic morphology, and (C) qPCR analysis for the neuronal markers NeuN (RBFOX3), MAP-2 (MAP2), class III β-tubulin (TuJ1; gene TUBB3), and synaptotagmin (SYT1), as indicated. NPC phenotype was defined by downregulation of the pluripotency marker Oct4 (POU5F1) and concomitant upregulation of nestin (NES). Expression was calculated using β-actin as an internal control. Error bars indicate SD based on 1 triplicate experiment.

Article Snippet: TaqMan qPCR (ThermoFisher) was used to confirm expression of the neuronal markers with the following TaqMan assays (ThermoFisher): NeuN (RBFOX3) 4331182-Hs01370654_m1); TuJ1 (Class III β-tubulin, TUBB3) 4331182-Hs00801390_s1; synaptotagmin (SYT1) 4331182-Hs00194572_m1; and MAP-2 (MAP2) 4331182-Hs00258900_m1, as described above.

Techniques: Expressing, Derivative Assay, Control, Generated, Microscopy, Marker