zm 241385  (Tocris)

Journal: Stem Cell Research & Therapy

Article Title: Molecular and functional characterization of GMP-manufactured neural stem cells and their extracellular vesicles for innovative therapeutic applications

doi: 10.1186/s13287-026-04904-x

Figure Lengend Snippet: hNSCs characterization. A Cumulative growth curve over 5 weeks of culture; B Clonal efficiency determined in three different hNSCs batches (CE1: clonal efficiency batch 1; CE2: clonal efficiency batch 2; CE3: clonal efficiency batch 3); C Representative images of hNSCs grown as neurospheres after 7 days of culture (magnification 10x); IHC performed on paraffin-embedded hNSCs neurospheres stained for Hematoxylin and Eosin, Ki-67, Olig2, GFAP and NSE (magnification 40x); D Expression of OCT4 and E KLF4. Representative flow cytometry histograms showing the expression levels of OCT4 and KLF4 in hNSCs. Specific antibody staining is shown in red, and isotype control in grey. The numbers indicate the percentage of positive cells relative to the isotype control; F Western blotting analysis of SOX2 in hNSC total cell lysate. THP-1 cells were used as negative control; β-actin was used as loading control. The images are representative of one out of three separate experiments; G Gene expression levels of ALPL, OCT4, KLF4, SOX2 were quantified using the 2 −ΔCt method, with GAPDH as the internal control. Data represent mean ± SD from three independent experiments

Article Snippet: In independent experiments, THP-1 cells were treated with 500 nmol/L ZM-241385 (4-(2-[7-A mino-2-(2-furyl) [ , , ]triazolo[2,3- a ] [ , , ]triazin-5-ylamino]ethyl)phenol, (Tocris, Bio-techne, Minneapolis, MN, USA) 1 h before treatments.

Techniques: Staining, Expressing, Flow Cytometry, Control, Western Blot, Negative Control, Gene Expression

Journal: Stem Cell Research & Therapy

Article Title: Molecular and functional characterization of GMP-manufactured neural stem cells and their extracellular vesicles for innovative therapeutic applications

doi: 10.1186/s13287-026-04904-x

Figure Lengend Snippet: Immunomodulatory effects of hNSC-EVs on THP-1 monocytic cells. A hNSC-EVs uptake by THP-1 cells. Cells were treated with DiL-labeled hNSC-EVs in red for 1 h and then exposed for further 1 h with or without LPS + ATP; phalloidin in green was used for actin filaments staining and nuclei in blue were counterstained with Hoechst 33,258. Upper images show the whole z-stack section (as reported CTRL z:12/23; EVs z:18/36; EVs + LPS + ATP z:17/35) with a 50 μm white scale bar. The bottom panels represent the zoomed orthogonal images (fuchsia box) from the z-stack acquisition with a 5 μm white scale bar; B Caspase-1 activation. THP-1 cells were pre-treated for 1 h with 1000 hNSC-EVs/cell, subsequently primed for 20 min with 10 µg/ml LPS and then activated for 40 min with 5 mmol/L ATP (LPS). In selected experiments ZM-241,385 (500 nmol/L) was added 1 h before hNSC-EVs treatment; cell lysates were immunoblotted for Caspase-1. The blots were re-probed with mouse anti-β-actin, to confirm equal loading. Representative western blots images are shown. Histogram on the right represents densitometric quantification and indicates the mean ± SD of at least n = 3 independent experiments performed with EVs isolated from different hNSCs batches

Article Snippet: In independent experiments, THP-1 cells were treated with 500 nmol/L ZM-241385 (4-(2-[7-A mino-2-(2-furyl) [ , , ]triazolo[2,3- a ] [ , , ]triazin-5-ylamino]ethyl)phenol, (Tocris, Bio-techne, Minneapolis, MN, USA) 1 h before treatments.

Techniques: Labeling, Staining, Activation Assay, Western Blot, Isolation