Journal: Pediatric research

Article Title: Pathophysiology of childhood polycystic kidney diseases: new insights into disease-specific therapy

doi: 10.1038/pr.2013.191

Figure Lengend Snippet: The Future: Therapies for Childhood Pkd

Article Snippet: SMAC Mimetics/ HDAC-i/TNF-α , ++/? , ++/? , ( ) ( ) .

Techniques:

Journal: Journal of molecular modeling

Article Title: Molecular Modeling in the Age of Clinical Genomics, The Enterprise of the Next Generation

doi: 10.1007/s00894-017-3258-3

Figure Lengend Snippet: Known structures of XIAP Structures highlighted in yellow were used for modeling the complete XIAP structure ( Figure 2B ) and those in red used to model XIAP-protein interactions ( ).

Article Snippet: 4EC4 , X-ray , 3.3 , A/B/C/D/E/F/G/J/K/L , 241–356 , BIR3 , SMAC MIMETIC (0O6) , .

Techniques: Binding Assay

Journal: bioRxiv

Article Title: Systematic genetic mapping of necroptosis identifies SLC39A7 as modulator of death receptor trafficking

doi: 10.1101/290718

Figure Lengend Snippet: ( a ) Cell viability of KMB7 wildtype and KBM7 FADD - cells treated for 24h with 100 ng/ml TNFα, 1 µM SMAC mimetic birinapant, 50 µM Nec-1s and 10 µM z-VAD-FMK as indicated. ( b ) Cell viability of KBM7 FADD - cells treated for 24h with 100 ng/ml TNFα, 10 µM SMAC mimetic or 100 ng/ml TNFα and 1 µM SMAC mimetic, and the indicated inhibitors. Cell viability was assessed using a luminescence-based readout for ATP (CellTiter Glo) and normalized to untreated control. Data represent mean value ± s.d. of two independent experiments performed in triplicates. ( c ) Schematic overview summarizing the genetic screens presented in this study.

Article Snippet: Interestingly, we found that treatment with the SMAC mimetic birinapant alone sufficed to induce necroptosis in KBM7 FADD - cells ( ).

Techniques:

Journal: bioRxiv

Article Title: Systematic genetic mapping of necroptosis identifies SLC39A7 as modulator of death receptor trafficking

doi: 10.1101/290718

Figure Lengend Snippet: ( a-c ), Circos plots of haploid genetic screens in KBM7 FADD - cells with necroptosis induction by 10 µM SMAC mimetic birinapant ( a ), 100 ng/ml TNFα ( b ), and 1 µM SMAC mimetic and 100 ng/ml TNFα combined ( c ). Each dot represents a mutagenized gene identified in the resistant cell population, dot size corresponds to the number of independent insertions identified for each gene and distance from center indicates the significance of enrichment compared to an unselected control data set. Hits with an adjusted p -value <10 -10 are labelled. ( d ) Bubble plot depicting the top hits over all three screens ranked according to adjusted p -value. Bubble size corresponds to the number of independent insertions identified and colour gradient reflects the significance of enrichment. ( e-f ), Multi-colour competition assay (MCA) of KBM7 FADD - SpCas9 cells transduced with a GFP marker (GFP + ) and sgRNAs targeting either SLC39A7 or RIPK1 ( e ), SP1 or TNFR2 ( f ), or Renilla luciferase ( sgRen ) as control, against cells transduced with sg Ren and an mCherry marker (mCherry + ). The cell populations were mixed at 1:1 ratio, treated with SMAC mimetic (1 µM) or TNFα (10 ng/ml), and analyzed after 14 days by flow cytometry. Data represent mean value ± s.d. of two independent experiments performed in duplicates, n.d. (not determined) indicates wells with no outgrowth.

Article Snippet: Interestingly, we found that treatment with the SMAC mimetic birinapant alone sufficed to induce necroptosis in KBM7 FADD - cells ( ).

Techniques: Competitive Binding Assay, Transduction, Marker, Luciferase, Flow Cytometry

Journal: bioRxiv

Article Title: Systematic genetic mapping of necroptosis identifies SLC39A7 as modulator of death receptor trafficking

doi: 10.1101/290718

Figure Lengend Snippet: ( a - b ) Circos plots of genome-scale SAM screens in KBM7 FADD - SAM cells with necroptosis induction by TNFα ( a ) or SMAC mimetic birinapant ( b ). For each stimulus, screens were performed at low and high concentrations (TNFα: 10 ng/ml (green) or 100 ng/ml (purple); birinapant: 0.1 µM (blue) or 1 µM (orange)). Screen analysis was performed by identifying differentially enriched sgRNAs using DESeq2 and then aggregating sgRNAs to genes using Gene Set Enrichment Analysis. Identified hits are ranked according to the adjusted p -value of enrichment (–log10(padj). Bubble size corresponds to the average log2 fold-change (aLFC) of enrichment, color indicates the number of significantly enriched sgRNAs. Screens were performed in duplicate except the high concentration of Birinipant in simplicate. (c-d) Cell viability in KBM7 FADD - SAM cells transduced with sgRNA targeting TNIP1, BIRC3 or Renilla luciferase (Ren). Cells were treated as indicated for 72h and viability was assessed using a luminescence-based readout for ATP (CellTiter Glo). Data represent mean value ± s.d. of two independent experiments performed in triplicates. ( e ) KBM7 FADD - CRISPR-SAM cells transduced with the specified sgRNAs were lysed and subjected to immunoblotting with the indicated antibodies.

Article Snippet: Interestingly, we found that treatment with the SMAC mimetic birinapant alone sufficed to induce necroptosis in KBM7 FADD - cells ( ).

Techniques: Concentration Assay, Transduction, Luciferase, CRISPR, Western Blot

Journal: Journal of carcinogenesis & mutagenesis

Article Title: Inhibitor of Apoptosis Proteins: Promising Targets for Cancer Therapy

doi: 10.4172/2157-2518.S14-004

Figure Lengend Snippet: Pre-clinical data where IAP inhibition sensitised to anti-cancer therapies.

Article Snippet: , Smac mimetc (BV6) , Radiotherapy , [ ] .

Techniques: Inhibition, shRNA

Journal: PLoS ONE

Article Title: EGFR-Targeted TRAIL and a Smac Mimetic Synergize to Overcome Apoptosis Resistance in KRAS Mutant Colorectal Cancer Cells

doi: 10.1371/journal.pone.0107165

Figure Lengend Snippet: (a–d) Caco-2tet Ras G12V cells were seeded into 3D cultures in the presence of doxycycline. (a) Three days later, cultures were left untreated or treated with 5 µM SM83 or 20 µM Z-VAD for 1 h prior to addition of 1 nM Db αEGFR -scTRAIL. Viability was measured 24 h later by MTT assay and normalized to the untreated control (n = 3). (b) 24 h after treatment, cells were fixed and stained for DNA strand breaks. Tunel-positive cells were counted (n = 2). (c) Three days post seeding, cultures were left untreated (ut) or treated with 5 µM SM83. Cells were recovered from the cultures 24 h later and lysates were analyzed by immunoblotting. Shown is one representative blot of two independent experiments. Tubulin was detected as a loading control. (d) Quantification of Western blots from (c). Protein levels were normalized to the corresponding tubulin control; levels in the untreated cultures were set as 1 (n = 2). (e, f) HCT-116 and LoVo cells were grown in 3D cultures for six days, and then fixed and stained for F-actin and DNA (DAPI) (scale bar: 20 µm) (top panels). Three days post seeding, cultures were left untreated or pretreated with 5 µM SM83 prior to addition of 0.05 nM Db αEGFR -scTRAIL. Viability was measured 24 h later by MTT assay and normalized to the untreated control (bottom panels) (n = 3).

Article Snippet: In the presence of doxycycline, these cells showed increased resistance to Db αEGFR -scTRAIL, associated with the elevated expression of the anti-apoptotic proteins cIAP2, Bcl-xL and Flip S . Co-treatment of cells with the Smac mimetic SM83 restored the Db αEGFR -scTRAIL-induced apoptotic response.

Techniques: MTT Assay, Control, Staining, TUNEL Assay, Western Blot