Journal: bioRxiv

Article Title: Global and single-cell proteomics view of the co-evolution between neural progenitors and breast cancer cells in a co-culture model

doi: 10.1101/2023.05.03.539050

Figure Lengend Snippet: a Sp8 confocal images showing the marker presence of INA, NeuN, DCX, Nestin, Ki67, NFH and TH in the XCL-1 DCXp-GFP cell line. Scale bar – 40 μm b Brightfield images displaying the morphological change in the H9inGFPhESCs upon differentiation into NPC and subsequent differentiation into midbrain dopaminergic neurons, immature neurons (after two weeks of differentiation) and maturing neurons (after five weeks of differentiation). Scale bar – 200 μm. c Selected protein abundances of neural markers following the differentiation steps of NPCs, midbrain neurons, immature neurons, maturing neurons, NPC after co-culture with MDA-MB-231 spheroids and with MCF7 spheroids.

Article Snippet: Following the whole mounting protocol, neural cells were stained with rabbit polyclonal anti-tyrosine hydroxylase antibody (Merck, AB152, 1:200 dilution), guinea pig polyclonal anti-DCX antibody (Merck, AB2253, 1:200 dilution), rabbit polyclonal anti-INA antibody (Merck, AB5354, 1:200 dilution), mouse monoclonal anti-NeuN antibody (Merck, MAB377, 1:200 dilution), mouse monoclonal anti-Nestin antibody (CST, 33475, 1:200 dilution), rabbit monoclonal anti-Ki-67 antibody (Epredia, RM9106S, 1:200 dilution), chicken polyclonal anti-NFH antibody (Merck, AB5539, 1:200 dilution).

Techniques: Marker, Co-Culture Assay

Journal: Cell stem cell

Article Title: Zika Virus Targets Glioblastoma Stem Cells through a SOX2-Integrin α v β 5 Axis

doi: 10.1016/j.stem.2019.11.016

Figure Lengend Snippet: (A) Representative images of mock- or ZIKV-infected BCOs stained with neuronal markers (CTIP2 and NeuN), a neural progenitor cell marker (SOX2), and DAPI. Scale bars, 100 μm. (B) Quantification of BCO size p.i. with ZIKV. Significance was assessed by two-tailed Student’s t test, and experiments were performed in two batches with 12 organoids per group per batch. (C) BCO size fold change of ZIKV- and mock-treated groups over a period of 1 month. (D) Quantification of SOX2+ cells in ZIKV- versus mock-infected groups. *p < 0.05 by two-tailed Student’s t test. (E) Quantification of CC3+ cells in ZIKV- versus mock-infected groups. *p < 0.05 by two-tailed Student’s t test. (F) Quantification of SATB2+ cells within MAP2+ cells in ZIKV- versus mock-infected groups. **p < 0.01 by two-tailed Student’s t test. (G) Quantification of GFAP+ cells in ZIKV- versus mock-infected groups. N.S., not significant by two-tailed Student’s t test. (H) Quantification of NeuN+ cells in ZIKV- versus mock-infected groups. N.S., not significant by two-tailed Student’s t test. (I) Quantification of CTIP2+ cells in ZIKV- versus mock-infected groups. N.S., not significant by two-tailed Student’s t test. (J) Bright-field images of engraftment of two patient-derived GSCs (387 and 3565) transduced with GFP into human BCOs over a time course. Scale bars, 1 mm. (K) Engrafted GSCs (GFP+) with normal BCO immunostained for integrin αvβ5 (red), GFP (green), and DAPI (blue). Scale bars, 200 μm. (L) Quantification of integrin αvβ5+ cells in normal BCOs or GSC-BCOs. Values represent mean ± SEM. n = 6. ****p < 0.0001 by two-tailed Student’s t test. (M) Representative images of GFP-labeled GSC-BCOs immunostained for integrin αvβ5 (red), GFP (green), and DAPI (blue). Scale bars, 100 μm. (N) Representative images of GFP-labeled GSC-BCOs immunostained for SOX2 (red), GFP (green), and DAPI (blue). Scale bars, 100 μm. (O) Images of GFP-labeled GSC-GFP BCOs 13 days p.i. with ZIKV. Scale bars, 1 mm. (P) Representative images of residual GSCs (green) and DAPI staining (blue) of GFP-labeled GSC-GFP BCOs cultured under mock conditions or with ZIKV for 2–4 weeks. Scale bars, 200 μm. The percentage of GFP+ cells among DAPI+ cells was quantified. Values represent mean ± SEM. n = 6. ****p < 0.0001 by two-way ANOVA. (Q) Representative immunostaining for integrin αvβ5 (red), GFP (green), ZIKV-E (white), and DAPI (blue) of GFP-labeled GSC-GFP BCOs mock- or ZIKV-infected for 2–4 weeks. Scale bars, 200 μm (left) and 100 μm (center). The percentage of ZIKV-E+ cells among integrin αvβ5 cells was quantified. Values represent mean ± SEM. n = 6. ****p < 0.0001 by two-tailed Student’s t test. (R) Representative images of 387 and 3565 GSC-BCOs with or without ZIKV, respectively, stained with SOX2, ZIKV-E, and DAPI. GFP shows the presence of GSCs (scale bars, 50 μm). ZIKV-E+, GFP+, and ZIKV-E+ cells among GFP+ cells were quantified by counting (two GSCs cell lines, two repeats, n = 12 organoids/group); *p < 0.05 by two-tailed Student’s t test. (S) Schematic of the experiment design. (T) Volcano plot showing differences between GSC-BCO ZIKV versus GSC-BCO mock. 113 genes were differentially expressed (greater than 1.5-fold) between these two groups (*p < 0.05). (U) Network analysis of genes differentially expressed upon ZIKV infection, represented as a bubble plot.

Article Snippet: Mouse monoclonal antibody to NeuN , PhosphoSolutions , Cat# 538-FOX3; RRID:AB_2560943.

Techniques: Infection, Staining, Marker, Two Tailed Test, Derivative Assay, Transduction, Labeling, Cell Culture, Immunostaining

Journal: Cell stem cell

Article Title: Zika Virus Targets Glioblastoma Stem Cells through a SOX2-Integrin α v β 5 Axis

doi: 10.1016/j.stem.2019.11.016

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Mouse monoclonal antibody to NeuN , PhosphoSolutions , Cat# 538-FOX3; RRID:AB_2560943.

Techniques: Control, Negative Control, Virus, Recombinant, In Vitro, Transfection, cDNA Synthesis, Plasmid Preparation, Imaging, TUNEL Assay, SYBR Green Assay, Reverse Transcription, Bradford Assay, shRNA, Software, Gene Expression, Flow Cytometry, Microscopy