sta-21  (Cedarlane)

Journal: Fluids and Barriers of the CNS

Article Title: Blood-brain barrier permeability increases with the differentiation of glioblastoma cells in vitro

doi: 10.1186/s12987-024-00590-0

Figure Lengend Snippet: IL-6 modulates BBB permeability via STAT3 activation in blood-brain barrier cells. (a) Schematic representation of IL-6/STAT3 signalling. (b) hCMEC/D3 cells, grown for 7 days up to confluence in Transwell insert, were left untreated (CTRL) or incubated in the lower chamber with the conditioned medium derived from 5-day culture of GBM cells of patient #2, as medium from differentiated/adherent cells (AC), alone or containing an anti-IL-6 neutralizing antibody (Nab, 1/400), or medium from neurospheres (NS), alone or containing rhIL-6 (200 pg/mL). Immunoblot of STAT3 and phospho(Tyr705)STAT3. The expression of GAPDH was used as control of equal protein loading. The figure is representative of one out of three experiments with similar results. (c) BBB cells were grown as in b . When indicated the inhibitor of STAT3, STA-21 (30 µM) was co-incubated. Immunoblot of STAT3 and phospho(Tyr705)STAT3. The expression of GAPDH was used as control of equal protein loading. The figure is representative of one out of three experiments with similar results. (d) Immunoblotting of the indicated proteins in BBB cells, incubated as in c . The expression of GAPDH was used as control of equal protein loading. The figure is representative of one out of three experiments with similar results. ZO1: zonula occludens 1; CLDN3: claudin-3; CLDN5: claudin-5. (e) Permeability assay on BBB cells treated as in c . 5 µM doxorubicin, 10 µM mitoxantrone or 2 µM dextran 70-fluorescein isothiocyanate were added for 3 h. The compounds recovered from the lower chamber were measured fluorometrically, in duplicates. Data were presented as mean ± SD ( n = 3 independent experiments). * p < 0.05, *** p < 0.001: AC/NS vs. CTRL; ° p < 0.05, °° p < 0.01: STA-21-treated AC vs. AC, IL-6-treated NS vs. NS; # p < 0.05; STA-21 + IL-6 treated NS vs. IL-6 treated NS

Article Snippet: When indicated, the STAT3 inhibitor STA-21 (CAS 28882-53-3, Santa Cruz Biotechnology Inc., Santa Cruz, CA; 30 μM) or the proteolysis targeting chimera (PROTAC) against STAT3 SD-36 (#HY-129602, MedChemExpress, Monmouth Junction, NJ; 1 μM) were added in the Transwell insert containing hCMEC/D3 cells.

Techniques: Permeability, Activation Assay, Incubation, Derivative Assay, Western Blot, Expressing, Control

Journal: PLoS ONE

Article Title: Interleukin-10 Inhibits Lipopolysaccharide Induced miR-155 Precursor Stability and Maturation

doi: 10.1371/journal.pone.0071336

Figure Lengend Snippet: (A) RAW264.7 cells were transfected with the c-fos promoter reporter and TK-Renilla, and were pretreated with DMSO or 30 µM STA-21 for 1 hour prior to IL-10 stimulation for 6 hours. Luciferase activity was measured and plotted as firefly/renilla ratio. (B) SCRMB and SHIP1 siRNA transduced cells were treated as except the cells were pretreated with DMSO or 30 µM STA-21 for 1 hour prior to stimulation. Expression levels of pri-miR-155 at 2 hours and miR-155 at 4 hours were measured by real time PCR and plotted relative to the LPS alone samples. Statistical significance between stimulation conditions was calculated by a two-way ANOVA test with a 95% confidence (** p <0.01, *** p <0.001, **** p <0.0001). Results were observed in at least two independent experiments.

Article Snippet: AQX-MN100 (Aquinox Pharmaceuticals, Vancouver, BC) was dissolved in ethanol, and STA-21 (Cedarlane Laboratories, Burlington, ON) was dissolved in dimethylsulfoxide (DMSO).

Techniques: Transfection, Luciferase, Activity Assay, Expressing, Real-time Polymerase Chain Reaction