Journal: iScience

Article Title: Tumor microenvironment mimicking 3D models unveil the multifaceted effects of SMAC mimetics

doi: 10.1016/j.isci.2023.106381

Figure Lengend Snippet: SM induces human dendritic cell maturation which is further enhanced by cancer cell necroptosis (A) Schematic representation of the in vitro co-culture assay. moDCs alone (B) or in co-culture with spheroids (tumor cells and hd-fibroblasts) (D and E) were treated with the compound conditions depicted. 72 h post-treatment, cells were harvested and analyzed via flow cytometry. (B) Relative proportion of activated vs non-activated moDC treated alone. (C) TSNE plot depicting islands of phenotypically similar cells based on CD45, CD11c, HLA-DR, CD86, CD83, and PDL1 as markers. Included are the moDCs treated in monoculture, as well as the different co-cultures. Analyzed cells separate into three subpopulations: activated moDC – blue, non-activated moDC - light gray, tumor cells - black. (D) Relative proportion of tumor and moDCs in the different co-cultures. (E) Relative proportion of activated vs non-activated moDCs in co-culture. Error bars, mean ± SD of triplicate independent wells for each condition of at least 2 independent experiments using different donors. ∗∗p value <0.01, by two-tailed t-test. TNFα: 0.1 ng/mL, SM: 1 μM, zVad: 5 μM. (F) Heatmap depicting cytokines measured in the supernatant of spheroids composed of tumor cells with or without CAFs, 72 h post-treatment. Data are averages from 3 independent wells and black squares indicate levels measured below the detection limit. SM: 0.25-1 μM; TNFα: 0.1 ng/mL, zVad: 20 μM.

Article Snippet: The human dermal fibroblasts (hd-fibroblasts - ATCC® PCS-201-010TM) were cultured in Fibroblast Basal Medium (ATCC PCS201030) supplemented 2% heat-inactivated FBS and Fibroblast Growth Kit-Serum Low (ATCC, #PCS-201-041) without HLL Supplement and rh EGF/TGTb1.

Techniques: In Vitro, Co-culture Assay, Co-Culture Assay, Flow Cytometry, Two Tailed Test

Journal: iScience

Article Title: Tumor microenvironment mimicking 3D models unveil the multifaceted effects of SMAC mimetics

doi: 10.1016/j.isci.2023.106381

Figure Lengend Snippet: SM modulates fibroblasts by downregulating myofibroblast-like CAF markers and upregulating pro-inflammatory soluble mediators CAF 2D monoculture was treated with increasing concentrations of SM or TGFβ. (A) Fold change in FAP and αSMA expression, analyzed via flow cytometry, 72 h post-treatment. (B) CAF confluence monitored overtime using IncuCyte. Error bars, mean ± SD of 3 independent experiments. (C) Heatmap depicting fold change in cytokine and chemokine concentration in the supernatant of treated CAFs (normalized to control). Results from 2 independent experiments. Black squares indicate levels measured below the detection limit.

Article Snippet: The human dermal fibroblasts (hd-fibroblasts - ATCC® PCS-201-010TM) were cultured in Fibroblast Basal Medium (ATCC PCS201030) supplemented 2% heat-inactivated FBS and Fibroblast Growth Kit-Serum Low (ATCC, #PCS-201-041) without HLL Supplement and rh EGF/TGTb1.

Techniques: Expressing, Flow Cytometry, Concentration Assay, Control

Journal: iScience

Article Title: Tumor microenvironment mimicking 3D models unveil the multifaceted effects of SMAC mimetics

doi: 10.1016/j.isci.2023.106381

Figure Lengend Snippet: Key resources table

Article Snippet: The human dermal fibroblasts (hd-fibroblasts - ATCC® PCS-201-010TM) were cultured in Fibroblast Basal Medium (ATCC PCS201030) supplemented 2% heat-inactivated FBS and Fibroblast Growth Kit-Serum Low (ATCC, #PCS-201-041) without HLL Supplement and rh EGF/TGTb1.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Software, Simple Western, High Content Screening

Journal: International Journal for Parasitology: Drugs and Drug Resistance

Article Title: In vitro activity, safety and in vivo efficacy of the novel bumped kinase inhibitor BKI-1748 in non-pregnant and pregnant mice experimentally infected with Neospora caninum tachyzoites and Toxoplasma gondii oocysts

doi: 10.1016/j.ijpddr.2021.05.001

Figure Lengend Snippet: Structure and in vitro activities of BKI-1748 against N. caninum (Nc-β-gal) and T. gondii (Tg-β-gal). (A) molecular structure of BKI-1748, MW = molecular weight; EC 50 = concentration of half maximal proliferation inhibition; *toxicity levels against human foreskin fibroblasts (HFF) and zebrafish embryos were determined previously [22]. (B,C) Dose-response curves for Neospora and Toxoplasma tachyzoites grown in HFF.

Article Snippet: Human foreskin fibroblasts (HFF; PCS-201-010TM) and BALB/c dermal fibroblasts (CELLNTEC AG, Bern, Switzerland) were maintained as described ( ).

Techniques: In Vitro, Molecular Weight, Concentration Assay, Inhibition

Journal: International Journal for Parasitology: Drugs and Drug Resistance

Article Title: In vitro activity, safety and in vivo efficacy of the novel bumped kinase inhibitor BKI-1748 in non-pregnant and pregnant mice experimentally infected with Neospora caninum tachyzoites and Toxoplasma gondii oocysts

doi: 10.1016/j.ijpddr.2021.05.001

Figure Lengend Snippet: Litter size, parasite burden, neonatal and postnatal mortality rates of N. caninum infected mice treated with BKI-1748.

Article Snippet: Human foreskin fibroblasts (HFF; PCS-201-010TM) and BALB/c dermal fibroblasts (CELLNTEC AG, Bern, Switzerland) were maintained as described ( ).

Techniques: Infection