Journal: Microarrays

Article Title: Time-Resolved Study of Nanoparticle Induced Apoptosis Using Microfabricated Single Cell Arrays

doi: 10.3390/microarrays5020008

Figure Lengend Snippet: Representative time traces of fluorescent signals monitored in the polarity Sensor for Viability and Apoptosis (pSIVA) ( A ) and propidium iodide (PI) ( B ) channels. A few typical traces are highlighted in green and red, respectively. The first apoptotic events are observed after 3 h whereas the majority of induced cell deaths occur after 20 h. The decrease in the pSIVA signal at late times can be attributed to bleaching of the fluorophores. Fluorescent signals are background corrected.

Article Snippet: Cells were stained with polarity Sensor for Viability and Apoptosis (pSIVA-IANBD, Novus Biologicals) [ , ] (the stock solution was diluted 1:50) and 1 μ M propidium iodide (Novus Biologicals).

Techniques:

Journal: Neuro-Oncology

Article Title: Multiplexed epigenetic memory editing using CRISPRoff sensitizes glioblastoma to chemotherapy

doi: 10.1093/neuonc/noaf055

Figure Lengend Snippet: CRISPRoff targeting of MGMT in primary GBM. (A) Western blots against MGMT protein in monoclonal (SF7996) or polyclonal (SF14259, SF14346, SF14590, SF14599) populations of patient-derived primary GBM cultures at the indicated number of days following delivery of CRISPRoff and the indicated sgRNA(s). (B) Crystal violet assay of cell viability 7 days following cell seeding and TMZ treatment (replenished every 3 days) at the indicated concentrations for SF7996 primary GBM. Quantification of cell growth (right). Scale bar = 1000 µm. (C) Propidium iodide cell cycle analysis of SF7996 cells treated with TMZ. P value = 2 tailed Student’s t -test for S-phase compared to sgScrambled 20 µM TMZ condition, n = 3 biological replicates. (D) Brightfield light micrographs of CRISPRoff-modified primary GBM cultures (SF14259, SF14346, SF14590, SF14599) 15–35 days following cell seeding, corresponding to the time at which vehicle-treated cells reached confluency. TMZ was replenished every 3 days during culture. Integrated intensity from live cells was quantified (right) for each cell type. (E) Apoptosis assay of primary GBM cultures 7 days following cell seeding and TMZ treatment. P value = 2 tailed Student’s t -test compared to cognate sgScrambled.

Article Snippet: Cell cycle analysis was performed by 70% ethanol fixation followed by propidium iodide staining (Cell Signaling Technology, cat 4087S) analyzed on the Attune NxT Flow Cytometer 48 hours after treatment with the drug.

Techniques: Western Blot, Derivative Assay, Crystal Violet Assay, Cell Cycle Assay, Modification, Apoptosis Assay