Novel indolylmaleimide acts as GSK-3β inhibitor in human neural progenitor cells

Quantitative and kinetic profile of Wnt/β-catenin signaling components during human neural progenitor cell differentiation

“…The construction of expression plasmid pCAGGS-S33Y containing a mouse mutant S33Y β-catenin sequence with a C-terminal HA-tag has been described previously [14,15].…”

“…pTCF-EGFP plasmids, in which EGFP protein expression is driven by a promoter containing six TCF binding sites [21], and pCAGGS-S33Y for expression of stabilized β-catenin [14,15] were cotransfected into cells using Amaxa Nucleofector technology (Amaxa, Gaithersburg, USA) according to the manufacturer's instructions. Twenty-four hours after transfection, GFP-positive cells were analyzed by FACS analysis.…”

“…To verify whether TCF-dependent transcription is inducible by the Wnt/β-catenin pathway in ReNcell VM, a pTCF-EGFP plasmid, in which an EGFP gene is driven by a promoter containing TCF binding sites, and a pCAGGS-S33Y plasmid for overexpression of stabilized β-catenin [14,15] were cotransfected into ReNcell VM cells and the number of GFP-positive cells was measured 24 h after transfection by flow cytometry. Results show that compared to the control group with only pTCF-EGFP transfection, the number of GFPpositive cells in the cotransfected group increases by about 5-fold to 6% of the total cell population (Fig.…”

“…However, little is known about the kinetic regulation of the Wnt signals during these processes although the specificity of signaling pathways rely on their temporal and spatial dynamics, especially of the pathway's downstream signaling proteins [12]. Our previous studies showed that the Wnt/β-catenin pathway and Wnt3a, a typical activator of the Wnt/β-catenin pathway [13], regulate neurogenesis of ReNcell VM cells in vitro [14,15]. Thus, in the present study, we investigate the kinetic and dynamic Wnt/β-catenin pathway during physiological differentiation of ReNcell VM cells, in order to elucidate the role of this pathway during hNPC differentiation.…”

Interference of a novel indolylmaleimide with microtubules induces mitotic arrest and apoptosis in human progenitor and cancer cells

“…On one hand GSK-3b maintains proliferation of cells while on the other hand inhibition of GSK-3b can increase differentiation of cells. In a previously conducted screening [9] we tested newly synthesised indolylmaleimide derivatives, based on the well described GSK-3b inhibitor SB-216763 (SB21, Fig. 1), for their potential to similarly act as GSK3b inhibitors and their ability to induce differentiation in human neural progenitor cells (hNPCs).…”