Project description:We identified GSK3 as a regulator of GBM cell survival using microarray analysis, small molecule and genetic inhibitors of GSK3 activity. Various molecular and genetic approaches were then employed to dissect out the molecular mechanisms responsible for GSK3 inhibition-induced cytotoxicity. Keywords: time course
Project description:HeLa cell extracts with or without GSK3 enzyme inhibition were assayed using protein microarrays in order to detect GSK3-dependent changes in protein polyubiquitination.
Project description:Here we used human cortical brain organoids to probe the longitudinal impact of GSK3 inhibition through multiple developmental stages. Chronic GSK3 inhibition increased the proliferation of neural progenitors and caused massive derangement of cortical tissue architecture. Cortical organoids were differentiated as previously described (Paşca et al., 2015, doi: 10.1038/nmeth.3415.).Chronic GSK3 inhibition was performed by adding CHIR99021 (Merck SML1046) to the medium at day 0 (1 microM) and kept throughout the differentiation process until reaching the respective collection timepoints (day 18, day 50, day 100).
Project description:HeLa cell extracts with or without GSK3 enzyme inhibition were assayed using protein microarrays in order to detect GSK3-dependent changes in protein polyubiquitination. HeLa lysates in triplicates were supplemented with ubiquitin and incubated on protein microarrays (ProtoArray 5.0; Invitrogen) in the presence or absence of the GSK3 inhibitor SB-216763. Polyubiquitination of the arrayed proteins was detected using specific antibodies. ProtoArray 5.0 contains over 9,000 full-length human proteins purified and arrayed in duplicate under native conditions to maximize functionality.
Project description:To compare transcriptional changes of germinal center B cells upon GSK3 inhibition, induced GC B cells were treated with GSK3 inhibitor, CHIR99021. 407 differentially expressing genes were identified
Project description:We explored the molecular impact of blocking GSK3 on the gene expression profile in human monocytes. GSK3 inhibition by CHIR-99021 redirected monocytes towards acquiring an anti-inflammatory gene profile similar to M-CSF-primed monocyte-derived macrophages.
Project description:Here we used human cortical brain organoids to probe the longitudinal impact of GSK3 inhibition through multiple developmental stages. Chronic GSK3 inhibition increased the proliferation of neural progenitors and caused massive derangement of cortical tissue architecture. Cortical organoids were differentiated as previously described (Paşca et al., 2015, doi: 10.1038/nmeth.3415.). Chronic GSK3 inhibition was performed by adding CHIR99021 (Merck SML1046) to the medium at day 0 (1 microM) and kept throughout the differentiation process until reaching the respective collection timepoints (day 50, day 100).
Project description:The remarkable plasticity of macrophages is tied to extracelular cues and their subsequently signaling pathways. The transcriptional factor MAFB is critical to prompt the aquisition of an anti-inflammatory phenotype in macrophages. As MAFB activity and stability is highly regulated by GSK3, we explored the molecular impact of blocking GSK3 on the gene expression profile of human alveolar macrophages (AMØ), whose development and gene expression profile is GM-CSF-dependent. GSK3 inhibition skewed the transcriptional profile of AMØ towards an anti-inflammatory phenotype.