Journal: International Journal of Molecular Sciences

Article Title: A Fast and Reliable Process to Fabricate Regenerated Silk Fibroin Solution from Degummed Silk in 4 Hours

doi: 10.3390/ijms221910565

Figure Lengend Snippet: Cytocompatibility of silk fibroin films: Left : XTT assay for cell viability and proliferation of L929 cells cultivated on films from silk fibroin for 24 h, 48 h, and 72 h, respectively. The mean cell metabolic activity and standard error of the mean are reported for three independent silk fibroin samples dissolved with ZnCl 2 in triplicates. Right : Representative fluorescence microscopy image of live-dead staining of L929 cells after 72 h cultivated on films from silk fibroin dissolved in ZnCl 2 .

Article Snippet: The XTT Cell Proliferation Assay Kit (Serva, Heidelberg, Germany) was used to quantitatively evaluate cell metabolic activity according to the manufacturer’s protocol after 24, 48, and 72 h, respectively.

Techniques: XTT Assay, Activity Assay, Fluorescence, Microscopy, Staining

Journal: bioRxiv

Article Title: Therapeutic reversal of prenatal pontine ID1 signaling in DIPG

doi: 10.1101/2021.05.10.443452

Figure Lengend Snippet: (A) Western blot (WB) confirming ID1 knockdown in DIPG007 cells. (B) WB depicting reduction in SPARCL1 expression along with decreased ID1 expression in ID1 -knockdown DIPG007 cells. (C) Effect of ID1 knockdown on invasion as measured by Matrigel-coated Boyden chamber assay. Images show invading cells stained with crystal violet. Each data point represents an individual image; **P < 0.01, unpaired parametric t-test. (D-E) Effect of ID1 knockdown on DIPG007 migration as measured by scratch assay, quantified as percent wound closure. Images show representative scratch at 0 and 24 hours outlined in dotted red line. Experiment was completed in triplicate and data points represent mean+/-SEM, **P < 0.01; images taken with Incucyte; area measured by ImageJ. (F) WB for ID1 and ACTB expression in DIPG007 and PPK cells treated with increasing concentrations of CBD or DMSO control. (G) Viability of DIPG007 and PPK cells treated with increasing concentrations of CBD (0.5-20µM) relative to DMSO-treated control. Experiment was completed in triplicate and data points represent mean+/-SEM. (H) DIPG007 cells were treated for 2 days with DMSO (control), 2.5µM or 5µM CBD and invasion was measured by Matrigel-coated Boyden chamber. Each data point represents an individual image, mean+/-SEM; ****P < 0.0001, unpaired parametric t-test. (I) Effect of CBD treatment (5-10µM) on DIPG007 migration as measured by scratch assay, quantified as percent wound closure. Experiment was completed in triplicate and data points represent mean+/-SEM, **P < 0.005, two-way ANOVA t-test. (J) Histogram showing increase in DCF (ROS) with increasing doses of CBD. (K) Production of ROS mediates the inhibitory activity of CBD through ID1. DIPG007 cells were treated for 72 hours with vehicle (DMSO) or different concentrations of CBD (10, 8, 6, 4, 2, 1 µM) in the presence and absence of 50 µM TOC. IC 50 was 2.3µM for CBD treatment alone and 10.34µM for CBD + TOC; ****P < 0.0001, two-way ANOVA t-test. Cell proliferation was measured using XTT assay.

Article Snippet: After 72 hours, in vitro cell viability was monitored by XTT Cell Proliferation Assay kit (Cayman Chemical).

Techniques: Western Blot, Expressing, Boyden Chamber Assay, Staining, Migration, Wound Healing Assay, Activity Assay, XTT Assay