Additional file 1 of Anti-TIM3 chimeric antigen receptor-natural killer cells from engineered induced pluripotent stem cells effectively target acute myeloid leukemia cells

Additional file 1: Fig. S1. Representative flow cytometric analysis of HSPC markers CD34, CD43, and CD45 in floating cells on day 16 of NK cell differentiation culture. Fig. S2. Analysis of TIM3 surface expression by flow cytometric analysis. Fig. S3. Western blot analysis of various intracellular signaling pathways in CARTIM3 NK-92 cells in response to recombinant human TIM3 (200 μg/mL) at 4 hours in comparison to WT NK-92 cells

Additional file 1 of Cardiac glycoside ouabain efficiently targets leukemic stem cell apoptotic machinery independent of cell differentiation status

Additional file 1: Table S1. Clinical characteristics of AML patient-derived primary cells. Table S2. Profiles of different AML subpopulations at the time of isolation and in culture. Fig. S1. Purity of CD34+ HSPCs isolated from human cord blood and G-CSF-mobilized peripheral blood. Fig. S2. TIM3 expression in LSCs and normal HSPCs. Fig. S3. Apoptosis is a major mode of cell death induced by cardiac glycosides in AML cells. Fig. S4. Percentage of CD15+ blasts in live (Annexin V−) cells upon ouabain treatment in study model 2. Fig. S5. Ouabain had minimal effect on normal PBMCs. Fig. S6. Ouabain induces caspase activation in AML cells. Fig. S7. Profiles of key apoptosis regulatory proteins in response to ouabain in AML HL-60 cells. Fig. S8. Ouabain induces a rapid loss of c-FLIPL and c-FLIPS in LSCs. Fig. S9. Small molecule inhibition of Mcl-1 similarly caused apoptosis in LSCs and LPCs

Appalachian Mountaintop Mining Particulate Matter Induces Neoplastic Transformation of Human Bronchial Epithelial Cells and Promotes Tumor Formation

Epidemiological studies suggest that living near mountaintop coal mining (MTM) activities is one of the contributing factors for high lung cancer incidence. The purpose of this study was to investigate the long-term carcinogenic potential of MTM particulate matter (PM_MTM) exposure on human bronchial epithelial cells. Our results show that chronic exposure (3 months) to noncytotoxic, physiological relevant concentration (1 μg/mL) of PM_MTM, but not control particle PM_CON, induced neoplastic transformation, accelerated cell proliferation, and enhanced cell migration of the exposed lung cells. Xenograft transplantation of the PM_MTM-exposed cells in mice caused no apparent tumor formation, but promoted tumor growth of human lung carcinoma H460 cells, suggesting the tumor-promoting effect of PM_MTM. Chronic exposure to the main inorganic chemical constituent of PM_MTM, molybdenum but not silica, similarly induced cell transformation and tumor promotion, suggesting the contribution of molybdenum, at least in part, in the PM_MTM effects. These results provide new evidence for the carcinogenic potential of PM_MTM and support further risk assessment and implementation of exposure control for PM_MTM

Induction of Stemlike Cells with Fibrogenic Properties by Carbon Nanotubes and Its Role in Fibrogenesis

We developed a three-dimensional fibroblastic nodule model for fibrogenicity testing of nanomaterials and investigated the role of fibroblast stemlike cells (FSCs) in the fibrogenic process. We showed that carbon nanotubes (CNTs) induced fibroblastic nodule formation in primary human lung fibroblast cultures resembling the fibroblastic foci in clinical fibrosis and promoted FSCs that are highly fibrogenic and a potential driving force of fibrogenesis. This study provides a predictive 3D model and mechanistic insight on CNT fibrogenesis

Ingenuity Pathways Analysis software output for the downstream components of human Bcl-2-interactome.

<p>Twenty molecules of downstream components were reported to date and were labeled in orange, while upstream molecules were labeled in green.</p

Bcl-2 and p53 expression in BEAS-2B, BEAS-Cr and H460 cells.

<p>(<i>A</i>) Cells were either left untreated or treated with Cr(VI) (20 µM) for 12 h. Cell lysates were prepared and analyzed for Bcl-2 and p53 by Western blotting. β-actin was used as a loading control. Densitometry was performed to determine the relative levels of Bcl-2 and p53 compared to β-actin. Representative blots are shown. (<i>B</i>) BEAS-Cr cells were transiently transfected with pcDNA (vector) or p53 plasmid, and Bcl-2 and p53 expression levels were determined by Western blotting. Values are means (± SD) (<i>n</i> = 4). *<i>P</i><0.05 versus BEAS-2B controls. ^#<i>P</i><0.05 versus vector-transfected control.</p

Apoptosis response to Cr(VI) treatment in BEAS-2B, BEAS-Cr, and H460 cells.

<p>(<i>A</i>) BEAS-2B, BEAS-Cr and H460 cells were treated with or without Cr(VI) (20 µM) for 12 h and apoptosis was determined by Hoechst 33342 assay. (<i>B</i>) and (<i>C</i>) Effect of Bcl-2 knockdown on Cr(VI)-induced apoptosis of the respective cell lines and their mutants. Values are means (± SD) (<i>n</i> = 4). *<i>P</i><0.05 versus non-treated control. ^#<i>P</i><0.05 versus Cr(VI)-treated passage-control BEAS-2B cells. **<i>P</i><0.05 versus Cr(VI)-treated respective BEAS-Cr and H460 cells.</p

Effect of Bcl-2 knockdown on cell invasion and migration of BEAS-Cr and H460 cells.

<p>(<i>A</i>) BEAS-2B, BEAS-Cr and H460 cells (1×10⁵ cells) were added to Transwell® inserts coated with Matrigel® and incubated for 24 h. Invading cells were stained and counted under a light microscope. Plots show relative invasion of BEAS-2B, BEAS-Cr and H460 cells. (<i>B</i>) Effect of Bcl-2 on cell invasion of the respective cell lines and their mutants. Experiments were repeated with the indicated cell lines and analyzed for cell invasion. (<i>C</i>) Confluent monolayers of BEAS-2B, BEAS-Cr and H460 cells were wounded, and the cells were allowed to migrate for 24 h. Wound space was visualized by light microscopy and analyzed by comparing the relative change in wound space as compared to control cell monolayers. (<i>D</i>) Effect of Bcl-2 on cell migration of the respective cell lines and their mutants. Cells were wounded and analyzed for cell migration over a 24 h period. (<i>E</i>) Representative micrographs of cells stained for invasion are shown. Values are means (± SD) (<i>n</i> = 4). *<i>P</i><0.05 versus passage-control BEAS-2B cells. ^#<i>P</i><0.05 versus the respective BEAS-Cr and H460 cells.</p

Effect of Bcl-2 knockdown on tumor-associated properties <i>in vivo</i>.

<p>(<i>A</i>) Mice were injected subcutaneously with 1×10⁶ passage-control BEAS-2B, BEAS-Cr, or shBcl-2 BEAS-Cr cells. Tumor formation was determined at 14 d post-injection. Representative photographs are shown. (<i>B</i>) Mice were similarly injected with control BEAS-2B, H460, or shBcl-2 H460 cells. Tumor formation and representative micrographs at 14 d post-injection are shown. Data are means (± SD) (<i>n</i> = 4). *<i>P</i><0.05 versus passage-control BEAS-2B cells. ^#<i>P</i><0.05 versus the respective BEAS-Cr and H460 cells.</p

Effect of Bcl-2 knockdown on angiogenic activity of BEAS-Cr and H460 cells.

<p>(<i>A</i>) BEAS-2B, BEAS-Cr, and H460 cell supernatants were incubated with HUVEC cells, and endothelial capillary tube formation was detected under a light microscope. The number of nodes formed by the tubes were scored and plotted. (<i>B</i>) Effect of Bcl-2 knockdown on angiogenic activity of the respective cell lines. Experiments were performed with the indicated cell lines and analyzed for endothelial tube formation as described. (<b>C</b>) Representative micrographs of tube formation are shown. Values are means (± SD) (<i>n</i> = 4). *<i>P</i><0.05 versus passage-control BEAS-2B cells. ^#<i>P</i><0.05 versus the respective BEAS-Cr and H460 cell supernatants.</p