Journal: Phytomedicine : international journal of phytotherapy and phytopharmacology

Article Title: Caffeic acid selectively eliminates teratogenic human-induced pluripotent stem cells via apoptotic cell death.

doi: 10.1016/j.phymed.2022.154144

Figure Lengend Snippet: Fig. 4. Selective elimination of iPSCs in a mixed population with hDFs or MPCs. (A) CellTracker red CMTPX dye-labeled iPSCs and CellTracker green CMFDA dye- labeled hDFs were co-cultured, treated with 50, 100, and 200 μM CAA for 24 h, and then observed under a fluorescence microscope. Scale bar = 100 μM. (B) Varying numbers of iPSCs were co-cultured with hDFs labeled with CellTracker green CMFDA dye in the presence or absence of 100 μM CAA. After 24 h, cells were washed with PBS, harvested, and analyzed by flow cytometry. (C) Un-labeled iPSCs were co-cultured with MPCs labeled with CellTracker green CMFDA dye, and then incubated with 50, 100, and 200 μM CAA for 24 h. Cells were harvested and flow cytometry data were acquired. Data are representative of three independent experiments. DMSO was treated as vehicle control.

Article Snippet: Analysis of selective elimination of iPSCs in the mixed cell population To examine selective killing of iPSCs by CAA, iPSCs were cultured on Matrigel-coated 12-well culture plates in mTeSR1 and then fluorescently labeled with CellTracker Red CMTPX (Thermo Scientific) at 37 ◦C for 45 min. After washing with PBS, hDFs labeled with CellTracker Green CMFDA were added and co-cultured in the presence or absence of CAA.

Techniques: Labeling, Cell Culture, Fluorescence, Microscopy, Flow Cytometry, Incubation, Control

Journal: Brain Sciences

Article Title: Methamphetamine and the Synthetic Cathinone 3,4-Methylenedioxypyrovalerone (MDPV) Produce Persistent Effects on Prefrontal and Striatal Microglial Morphology and Neuroimmune Signaling Following Repeated Binge-like Intake in Male and Female Rats

doi: 10.3390/brainsci14050435

Figure Lengend Snippet: ( A ) Tiled confocal images of a representative coronal hemisphere stained for Iba1. ( B ) Compared to saline controls (n = 5–9 per group), decreased densities of Iba1+ cells were observed in the PFC of male and female animals self-administering METH (n = 5–7 per group). * p < 0.05 vs. saline. ( C ) Left , representative tissue section showing outlines of nuclei (red) stained by the far-red nuclear stain RedDot2, and TUNEL staining (green). Arrows highlight TUNEL+ nuclei. Middle and right , immunostaining of the PFC and DStr for the apoptosis marker cleaved caspase 3 (CC3, green) and microglial marker Iba1 (red), counterstained with nuclear marker RedDot2 (blue). Arrows denote co-localization of all three signals. ( D ) Quantification of TUNEL+ cells in the PFC and DStr of male and female rats self-administering METH (n = 5–7 per group), MDPV (n = 6–8 per group), or saline (n = 4–9 per group). * p < 0.05 vs. corresponding saline groups. ( E ) Quantification of co-localized CC3, Iba1, and nuclei in the PFC and DStr of male and female rats self-administering METH (n = 4–6 per group), MDPV (n = 4–6 per group), or saline (n = 5–6 per group). * p < 0.05 vs. corresponding saline groups. All data are shown as mean ± SEM. Open circles represent data values from individual animals.

Article Snippet: Sections were then incubated with a 1× solution of the far-red fluorescent nuclear stain RedDot2 (Biotium, Fremont, CA, USA) for 30 min; washed 1 × 10 min in PBS; coverslipped; sealed; and stored at 4 °C, protected from light, until analysis.

Techniques: Staining, Saline, TUNEL Assay, Immunostaining, Marker

Journal: bioRxiv

Article Title: p53 deletion rescues lethal microcephaly in a mouse model with neural stem cell abscission defects

doi: 10.1101/272393

Figure Lengend Snippet: A. Examples of E12.5 dissociated cortical NSCs at stages of mitosis and cytokinesis, labeled with PH3 (red) and AurkB (green). B. The mean percentages (+/- s.e.m.) of cycling NSCs(Ki67+) in mitosis or midbody stage are slightly reduced in Kif20b-/- cultures. C. Categorizing the dividing NSCs from B, there is a small increase of Kif20b-/- NSCs in anaphase/early telophase. For B and C, n = 3 coverslips from 3 embryos each, with 1499 control and 1421 Kif20b-/- Ki67+ cells. * p ≤ 0.05, Student’s t-test for (B), Chi Square and Fisher’s exact test for (C). All cultures dissociated from E12.5 cortices and fixed after 24 hours. Scale bar, 5 μ m.

Article Snippet: Antibodies used in this analysis were rabbit polyclonal anti-human cleaved-caspase 3 (Cell-Signaling 9661S, 1:250), mouse monoclonal anti-rat beta-III-tubulin (Tuj1) (BioLegend 801201, 1:500), rat monoclonal anti-mouse Tbr2 (ebioscience 14-4875, 1:200), rabbit polyclonal anti-mouse Pax6 (BioLegend PRB-278P, 1:200), Aurora B kinase mouse monoclonal anti-rat (BD Biosciences 611082, 1:300), rabbit polyclonal anti-human alpha-tubulin (Thermo Scientific RB-9281-P0, 1:300), rat monoclonal alpha-tubulin (Novus Biologicals NB600-506, 1:300), rabbit monoclonal anti-human PH3 (Cell Signaling 3458, 1:200), chicken polyclonal anti-mouse Nestin (Aves Labs NES, 1:600), rat monoclonal anti-human Ki67 (eBioscience 14-5698, 1:100), mouse monoclonal anti-human p53 (Millipore 05-224, 1:250), rabbit polyclonal anti-mouse p53 (Leica Biosystems NCL-L-p53-CM5p, 1:500), rabbit polyclonal anti-human Beta-Catenin (Sigma-Aldrich SAB4500545 1:1000), and polyclonal rabbit anti Zo-1 (rabbit, Invitrogen 61-7300,1:50).

Techniques: Labeling