Journal: Clinical Science (London, England : 1979)

Article Title: New insights into the pathophysiology and therapeutic targets of asthma and comorbid chronic rhinosinusitis with or without nasal polyposis

doi: 10.1042/CS20190281

Figure Lengend Snippet: Targeting immune mechanisms and therapeutic options in T2 and non-T2 inflammation

Article Snippet: , Reactive aldehyde species (RASP) , RASP inhibitor (small molecule) , ADX-629 , Aldeyra , Asthma , 2.

Techniques: Binding Assay, Recombinant, Variant Assay

Journal: Clinical Science (London, England : 1979)

Article Title: New insights into the pathophysiology and therapeutic targets of asthma and comorbid chronic rhinosinusitis with or without nasal polyposis

doi: 10.1042/CS20190281

Figure Lengend Snippet: Most of the current monoclonal antibodies and inhibitors target type 2 inflammation (registered biologics in blue letters). Anti – IgE therapy with omalizumab and ligelizumab is already very effective and safe approach to treat patients with difficult asthma. Similarly, anti-IL-5 monoclonal antibodies, particularly mepolizumab, represent an effective therapy of eosinophilic inflammation. Type 2 inflammation can be downregulated also by dupilumab targeting common subunit shared by IL-4 and IL-13 receptors. Recently, tepelezumab directed against thymic stromal lymphopoietin (TSLP) represents another perspective monoclonal antibody modulating type 2 inflammation with another drugs against alarmins IL-33 and IL-25 in clinical studies. Also the inhibitors of CRTH2 (receptor for PDG2) such as fevipiprant are in clinical studies. In non-type 2 inflammation, no biologics are registered for the therapy of asthma with monoclonal antibodies inhibiting IL-17/IL-23 pathway without sufficient clinical effects, so far. In the effector phases of allergic inflammation, antihistamines and leukotrienes are used for decades with emerging inhibitors of tryptase or reactive aldehyde species (RASP) and monoclonal antibodies blocking pro-inflammatory cytokines IL-6, IL-1 and IL-31 in clinical studies. In the case of anti-TNF therapy, the risk potential adverse effects prevail over benefits in asthma therapy. Finally, chemokine receptor inhibitors may represent another tool to block recruitment of immune cells in allergic inflammation. Created with BioRender Sofware (Ref.n.AB25BY6SZW).

Article Snippet: , Reactive aldehyde species (RASP) , RASP inhibitor (small molecule) , ADX-629 , Aldeyra , Asthma , 2.

Techniques: Blocking Assay

Journal: Oncotarget

Article Title: Downregulation of Bmi1 in breast cancer stem cells suppresses tumor growth and proliferation

doi: 10.18632/oncotarget.16317

Figure Lengend Snippet: (A) Cells treated with PTC 209 and FMMC 419II cells stably transfected with Bmi1 shRNA plasmid show a decrease in Bmi1 mRNA expression. (B) Purified mRNA from the cells was reverse transcribed into cDNA and then analyzed for Bmi1 mRNA expression with quantitative PCR using TaqMan gene expression assays. The fold difference in expression between control samples and the PTC 209 treated of the Bmi1 shRNA transfected samples was calculated using the average of the Ct (threshold cycle) per group, relative to the expression of the internal control gene GAPDH . (C) FMMC cells were treated with PTC 209 (2 μM and 5 μM) for 24 hours and the expression of Bmi1 protein was detected with western blot. (D) PTC 209 treatment decreased Bmi1 protein expression in FMMC cells. Results are represented as mean ± S.E.M., *P <0.01, ***P <0.005.

Article Snippet: For treatment with the Bmi1 small molecule inhibitor PTC 209 (Xcess Biosciences, San Diego, CA), cells were trypsinized, washed, and seeded into 6-well plates at around 30% confluency.

Techniques: Stable Transfection, Transfection, shRNA, Plasmid Preparation, Expressing, Purification, Reverse Transcription, Real-time Polymerase Chain Reaction, Gene Expression, Control, Western Blot

Journal: Oncotarget

Article Title: Downregulation of Bmi1 in breast cancer stem cells suppresses tumor growth and proliferation

doi: 10.18632/oncotarget.16317

Figure Lengend Snippet: (A) Untreated (control) FMMC 419II cells. (B) Cells treated with 2 μM PTC 209. (C) Cells treated with 5 μM PTC 209. (D) Results of PTC 209 treatment shown as bar graphs. There is a G0/G1 cell cycle arrest in FMMC 419II cells treated with PTC 209 when compared to untreated cells. Similarly, FMMC 419II cells that have been transfected with a Bmi1 shRNA show a G1 arrest. (E) Bar graphs of cell cycle profiles for FMMC 419II cells from control (F) , colony 2 (G) , colony 4 (H) , and colony 5 (I) . Cells stained with PI/RNAse staining buffer were run on a FACSAria flow cytometer and cell cycle progression was analyzed and quantified (D, E) using FlowJo.

Article Snippet: For treatment with the Bmi1 small molecule inhibitor PTC 209 (Xcess Biosciences, San Diego, CA), cells were trypsinized, washed, and seeded into 6-well plates at around 30% confluency.

Techniques: Control, Transfection, shRNA, Staining, Flow Cytometry

Journal: Oncotarget

Article Title: Downregulation of Bmi1 in breast cancer stem cells suppresses tumor growth and proliferation

doi: 10.18632/oncotarget.16317

Figure Lengend Snippet: Flow cytometry analysis of CD49f and CD24 expression for FMCC 419II cells treated with 2 μM PTC 209 versus Bmi1 shRNA transfected colonies 4 and 5 were carried out (A) , and the fluorescent intensities were quantitated (B) .

Article Snippet: For treatment with the Bmi1 small molecule inhibitor PTC 209 (Xcess Biosciences, San Diego, CA), cells were trypsinized, washed, and seeded into 6-well plates at around 30% confluency.

Techniques: Flow Cytometry, Expressing, shRNA, Transfection