Journal: bioRxiv

Article Title: Combining Scalable Organ Chip Platform with Deep Learning-Based Imaging Analysis for Cancer Therapeutic Screening

doi: 10.1101/2024.06.10.598272

Figure Lengend Snippet: A) Schematic of a standardized workflow for high-throughput screening on OC Plex-32 cancer models to measure drug effect B, C) Viability results for chips treated with Gemcitabine, Gefitinib, and Osimertinib for 48 hours B) by CellTiter-Glo ATP assay and C) by imaging assay (n=4). Imaging metrics utilized for dose response curves establishment were selected based on the Z’-factor ranking. The left graph shows the ratio of the area of viable cells to the area of all cells per chip. The right graphs demonstrate the area of viable cells. Data were normalized to the value of the vehicle controls in all graphs in B and C . D) Representative images of chips treated with Osimertinib at 0.01-100 μM for 48 hours and then stained with Hoechst33342 and EthD-2 for viability assessment. The right two image columns are showing the zoomed-in view of the region in the corresponding gray panes in the same row E) Correlation graphs comparing imaging results to CellTiter-Glo result for drug effect evaluation. Each dot represents one chip.

Article Snippet: The compounds used in these studies, Gefitinib (Medchem Express, HY-50895), Gemcitabine (MedChem Express, HY-17026), Osimertinib (HY-15722), and Savolitinib (HY-15959), Paclitaxel (HY-B0015), Regorafenib (HY-10331), Pemetrexed (HY-10820) were individually prepared in sterile DMSO as 1000x stocks (0.01, 0.1, 1, 10, 100 mM), aliquoted, and stored at -80 °C.

Techniques: High Throughput Screening Assay, ATP Assay, Imaging, Staining

Journal: Science Advances

Article Title: SETDB1 modulates the TGFβ response in Duchenne muscular dystrophy myotubes

doi: 10.1126/sciadv.adj8042

Figure Lengend Snippet: Antibodies list.

Article Snippet: SETDB1 (mouse) , Thermo Fisher Scientific, MA5-15722 , Immunofluorescence (1:200).

Techniques: Immunofluorescence, Western Blot

Journal: Molecular cancer research : MCR

Article Title: Aberrant induction of a mesenchymal/stem-cell program engages senescence in normal mammary epithelial cells.

doi: 10.1158/1541-7786.MCR-19-1181

Figure Lengend Snippet: a, Western analysis of SMAD2, SMAD3, and ACTIN (loading control) levels in SKD-HMEC expressing shRNAs targeting GFP (shGFP), SMAD2 (shSMAD2), or SMAD3 (shSMAD3). b-f, SKD-HMEC expressing shRNAs targeting GFP (shGFP), SMAD2 (shSMAD2), or SMAD3 (shSMAD3) were left untreated or treated with recombinant OSM [10 ng/mL] for 10 days and then subjected to the following assays. b, Relative growth assays of shGFP-, shSMAD2-, and shSMAD3-expressing HMEC grown in the presence (+) and absence (−) of OSM. Mean ± SEM (error bars) of all triplicates are presented for each sample. c, Senescence-associated β-galactosidase (SA-β-Gal) activity staining (blue) in untreated shGFP-, shSMAD2-, and shSMAD3-expressing HMEC and cells treated with OSM. Scale bar, 150 μm. d, Flow cytometry of CD24 and CD44 surface-profile expression in shGFP-, shSMAD2-, and shSMAD3-expressing SKD-HMEC grown in the presence (+) and absence (−) of OSM. CD44LO cells are shown as red and CD44HI cells as blue. Black numbers represent the percent of total cell population that are CD44HI. e, qRT-PCR analysis of cells grown in the presence (+) and absence (−) of OSM using primers targeting CD44s (SC gene; left) and SNAI1 (EMT gene; right) f, Following treatment with OSM [10 ng/mL] for 10 days, mRNA was harvested from shGFP-, shSMAD2-, or shSMAD3-expressing SKD-HMEC and subjected to a targeted Human EMT qRT-PCR profiler array. Heat map displaying non-supervised hierarchical clustering of fold-regulation gene expression data between OSM-treated cells relative to untreated shGFP-cells with the brightest green representing the smallest value, brightest red the highest value, and black the average magnitude of gene expression. Error bars indicate mean ± SEM of three independent experiments. Statistically significant differences are indicated with * when P < 0.05, ** when P < 0.01, *** when P < 0.001, and **** when P < 0.0001. Differences with a P value > 0.05 were considered non-significant and indicated with ns.

Article Snippet: 15 , 16 , 20 The retroviral plasmids pRetroSUPER-Puro shSMAD2 and pRetroSUPER-Puro shSMAD3 (Addgene plasmid #s 15722 and 15726) were gifts from and deposited to Addgene by Dr. Joan Massague (Memorial Sloan-Kettering Cancer Center); and pRetroSUPER-Puro shGFP was obtained from Dr. Yosef Shiloh (Department of Neurobiology, Tel Aviv University, Tel Aviv, Israel).

Techniques: Western Blot, Expressing, Recombinant, Activity Assay, Staining, Flow Cytometry, Quantitative RT-PCR