Journal: bioRxiv

Article Title: Notch1 regulates breast cancer stem cell function via a non-canonical cleavage-independent pathway

doi: 10.1101/2020.02.28.970764

Figure Lengend Snippet: Immunoblot analysis of: (A) Tsp-1, c-myc and actin expression in BPE cells in the presence and absence of ectopic expression of SV40 Large T antigen (LT) (BPLE), and HRasV12 (BPLER); (B) Myc, p53 and actin expression in wild type BPE cells transduced with vector controls pLKO.1-puro (V1), pMKO-neo (V2), pLKO.1shp53 (sh1) and pMKOneoshp53 (sh2); (C) Myc, p53 and actin expression in normal HME, normal human bronchial epithelial cells (NHBE) transduced with vector controls pLKO.1-puro (V1), pMKO-neo (V2), pLKO.1shp53 (sh1) and pMKOneoshp53 (sh2); (D) Myc, p53 and actin expression in BPE1, HME, and NHBE cells that were transiently transfected with pCMVneo or pCMVneo-p53 (p53); (E) Myc, p53 and actin expression in HME and BPE3 cells that were untreated (-) or treated with 25μM Nutlin-3: (F) Myc, p53 and actin expression in BPE and HME cells that were transiently transfected with pCMVneo, pCMVneo-p53 (wt) or pCMVneo-p53R175H; (G) Growth curve of HME, BPE, MCF-7 and MDA-MB-231 (231) cells that were untreated (black line) or treated with 25μM Nutlin-3/day for 5 days (Grey line); (H) Western blot of Myc, p53 and actin expression in A549 lung cancer cells that were untreated (-) or treated with 25μM Nutlin-3; (I) Western blot of Myc, p53 and actin expression in LNCaP prostate cancer cells that were untreated (-) or treated with 25μM Nutlin-3; (J) Western blot of Myc, p53 and actin expression in HCT116 colon cancer cells that were untreated (-) or treated with 25μM Nutlin-3.

Article Snippet: Human renal and lung (bronchial) epithelial cells (Lonza, Walkersville, MD) were immortalized via transduction of the retroviral vector pBabeHygro hTERT.

Techniques: Western Blot, Expressing, Transduction, Plasmid Preparation, Transfection

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Bacillus anthracis co-opts nitric oxide and host serum albumin for pathogenicity in hypoxic conditions

doi: 10.3389/fcimb.2013.00016

Figure Lengend Snippet: Supplementation of complete DMEM/F12 culture medium with BSA increases the toxicity of Sups of dSterne cultures grown in microaerobic conditions for 24 h. HSAECs were exposed to Sups for 2 h. Untreated cells served as control. OD 600 of Sups was measured in 96-well plate reader (200 μl per well). Error bars indicate 95% confidence intervals.

Article Snippet: Primary human small-airway lung epithelial cells (HSAECs) were from Cambrex, Inc., MD.

Techniques: Control

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Bacillus anthracis co-opts nitric oxide and host serum albumin for pathogenicity in hypoxic conditions

doi: 10.3389/fcimb.2013.00016

Figure Lengend Snippet: Antioxidant treatment of Sups protects HSAECs. The dSterne Sups grown in complete DMEM/F12 were incubated with 5 mM DTT for 30 min (A) or indicated concentrations of cysteine for 1 h (B) . HSAECs were exposed to the treated Sups for 2 h and the viability of cells was tested using Alamar Blue. In control experiments, addition of Cys had no effect on viability of untreated cells, and no cytoprotection was detected with other L - amino acids tested (Val, Leu, Ala) (not shown). Error bars indicate 95% confidence intervals.

Article Snippet: Primary human small-airway lung epithelial cells (HSAECs) were from Cambrex, Inc., MD.

Techniques: Incubation, Control

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Bacillus anthracis co-opts nitric oxide and host serum albumin for pathogenicity in hypoxic conditions

doi: 10.3389/fcimb.2013.00016

Figure Lengend Snippet: Inhibition of baNOS with L-NAME decreases toxicity of Sups to HSAECs. Static cultures of dSterne strain were grown in complete DMEM/F12 as described in Materials and Methods with or without the indicated concentrations of L-NAME. The table shows the ODs and pHs of 24-h cultures. The Sups were added for 2 h to HSAECs pretreated or not with the indicated concentrations of L-NAME for 1 h, and the viability of the cells was assayed with Alamar Blue. Controls included the culture medium with L-NAME without bacteria. All tested samples were titrated with HCl to the pH 5.1 of Sups without L-NAME. Error bars indicate 95% confidence intervals.

Article Snippet: Primary human small-airway lung epithelial cells (HSAECs) were from Cambrex, Inc., MD.

Techniques: Inhibition, Bacteria

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Bacillus anthracis co-opts nitric oxide and host serum albumin for pathogenicity in hypoxic conditions

doi: 10.3389/fcimb.2013.00016

Figure Lengend Snippet: Nitrosylated BSA (NO-BSA) is not acutely toxic. HSAECS were incubated with NO-BSA at indicated concentrations and pH for 2 h in DMEM/F12 medium, and the viability of HSAECs was tested using Alamar Blue. Error bars indicate 95% confidence intervals.

Article Snippet: Primary human small-airway lung epithelial cells (HSAECs) were from Cambrex, Inc., MD.

Techniques: Incubation

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Bacillus anthracis co-opts nitric oxide and host serum albumin for pathogenicity in hypoxic conditions

doi: 10.3389/fcimb.2013.00016

Figure Lengend Snippet: Blocking of free SH groups of BSA does not reduce the toxicity of dSterne Sup grown in the presence of BSA. The protein was dissolved in 10 ml of PBS, treated with 5 mM DTT for 30 min, and then dialyzed against two changes of 1 l of PBS for 24 h. The reduced BSA was then modified by NEM (125 μM) for 1 h at 37°C and dialyzed against two changes of 1 l of PBS overnight. The modified and mock-treated BSA was used to supplement the DMEM/F12 medium at 2 mg/ml. Bacterial cultures were grown for 24 h and Sups were tested for toxicity using HSAECs and the Alamar Blue viability assay. Error bars indicate 95% confidence intervals.

Article Snippet: Primary human small-airway lung epithelial cells (HSAECs) were from Cambrex, Inc., MD.

Techniques: Blocking Assay, Modification, Viability Assay

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Bacillus anthracis co-opts nitric oxide and host serum albumin for pathogenicity in hypoxic conditions

doi: 10.3389/fcimb.2013.00016

Figure Lengend Snippet: Permanganate treatment reduces the toxicity of dSterne Sup. The indicated concentrations of KMnO 4 were added for 1 h to the Sup grown in CSFM for 24 h, and toxicity of the treated Sup was tested after incubation with HSAECs for 2 h. Error bars indicate 95% confidence intervals.

Article Snippet: Primary human small-airway lung epithelial cells (HSAECs) were from Cambrex, Inc., MD.

Techniques: Incubation

Journal: Oncology Letters

Article Title: Antitumor activity of nimotuzumab in combination with cisplatin in lung cancer cell line A549 in vitro

doi: 10.3892/ol.2018.7923

Figure Lengend Snippet: Cyclin D1 expression in A549 cells in groups A-E following treatment for 48 h. Lanes: A, nimotuzumab; B, cisplatin; C, nimotuzumab followed by cisplatin; D, nimotuzumab and cisplatin simultaneously; and E, untreated control.

Article Snippet: Cell culture The A549 human lung adenocarcinoma epithelial cell line (supplied by the Central Laboratory of Qinhuangdao No. 1 People's Hospital) was cultured in Dulbecco's modified Eagle's medium (Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) containing 10% fetal bovine serum (FBS; Zhejiang Tianhang Biotechnology Co., Ltd., Huzhou, China), 1 U/ml penicillin and 1 mg/ml streptomycin at 37°C, with 5% CO 2 and 95% humidity.

Techniques: Expressing, Control

Journal: Toxicology letters

Article Title: Mitochondrial dysfunction is associated with Miro1 reduction in lung epithelial cells by cigarette smoke

doi: 10.1016/j.toxlet.2019.09.022

Figure Lengend Snippet: CSE reduces mitochondrial function as measured by oxygen consumption rate (OCR) an indicator of oxidative phosphorylation (OXPHOS) in human lung epithelial cells. BEAS2B, NHBE and DHBE cells were treated with CSE (0.5–1.0%) for 1–2 hrs or 24 h. Mitochondrial oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) was evaluated by using a Seahorse XFp analyzer in (A–B) BEAS2B, (C–D) NHBE and DHBE cells. CS-induced suppression of basal OCR, ECAR, maximal respiration, proton leak, ATP production and spare capacity. Data are mean ± SEM, n = 2–3 per group. *P < 0.05, **P < 0.01, vs respective controls.

Article Snippet: Cell culture and treatments Human small airway epithelial cells (SAEC), normal human bronchial epithelial cells (NHBE) from healthy non-smoking subjects and diseased human bronchial epithelial cells (DHBE) from patients with COPD were obtained from Lonza (Walkersville, MD), and grown as per the recommendations of the supplier in appropriate primary cell culture growth media.

Techniques: Phospho-proteomics