Journal: Microbiology Spectrum

Article Title: Epitranscriptomic N 6 -Methyladenosine Profile of SARS-CoV-2-Infected Human Lung Epithelial Cells

doi: 10.1128/spectrum.03943-22

Figure Lengend Snippet: SARS-CoV-2 infection of A549-hACE2 cells. A549-hACE2 cells were infected with SARS-CoV-2 (strain USA-WA1/2020) at an MOI of 1 for 24 h. (A) Immunofluorescent staining was performed to visualize infected cells by the presence of SARS-CoV-2 nucleocapsid (green). Nuclei of cells are stained with DAPI (blue). (B) SARS-CoV-2 spike RNA in infected cells ( n = 3, biological triplicate) was quantified by RT-qPCR. ND, not detected.

Article Snippet: A549-hACE2 lung carcinoma cells expressing the human ACE2 protein (Invivogen) were maintained in DMEM with 4.5 g/L glucose and 2 mM l -glutamine (Gibco), 10% heat-inactivated fetal bovine serum (FBS; R&D Systems), 100 U/mL penicillin, 100 μg/mL streptomycin (Gibco), and 0.5 μg/mL puromycin (Sigma).

Techniques: Infection, Staining, Quantitative RT-PCR

Journal: Microbiology Spectrum

Article Title: Epitranscriptomic N 6 -Methyladenosine Profile of SARS-CoV-2-Infected Human Lung Epithelial Cells

doi: 10.1128/spectrum.03943-22

Figure Lengend Snippet: Epitranscriptomic m 6 A microarray of SARS-CoV-2-infected A549-hACE2 cells. (A) Schematic overview of the method. Total cellular RNA from each sample (SARS-CoV-2-infected and mock-infected controls, biological triplicate, n = 3 each group) was used for immunoprecipitation using an m 6 A-specific antibody. Methylated and unmethylated RNA fractions were fluorescently labeled (Cy3 or Cy5) prior to array hybridization (refer to Materials and Methods for details). (B) Volcano plot of transcripts containing higher (red) and lower (blue) levels of m 6 A modification in infected cells compared to mock-infected control cells. The miRNA precursor (pre-mir-4486) with the most significant m 6 A change is labeled.

Article Snippet: A549-hACE2 lung carcinoma cells expressing the human ACE2 protein (Invivogen) were maintained in DMEM with 4.5 g/L glucose and 2 mM l -glutamine (Gibco), 10% heat-inactivated fetal bovine serum (FBS; R&D Systems), 100 U/mL penicillin, 100 μg/mL streptomycin (Gibco), and 0.5 μg/mL puromycin (Sigma).

Techniques: Microarray, Infection, Immunoprecipitation, Methylation, Labeling, Hybridization, Modification, Control

Journal: Microbiology Spectrum

Article Title: Epitranscriptomic N 6 -Methyladenosine Profile of SARS-CoV-2-Infected Human Lung Epithelial Cells

doi: 10.1128/spectrum.03943-22

Figure Lengend Snippet: Summary of differentially modified transcripts by type in SARS-CoV-2-infected A549-hACE2 cells versus mock-infected control cells ^a

Article Snippet: A549-hACE2 lung carcinoma cells expressing the human ACE2 protein (Invivogen) were maintained in DMEM with 4.5 g/L glucose and 2 mM l -glutamine (Gibco), 10% heat-inactivated fetal bovine serum (FBS; R&D Systems), 100 U/mL penicillin, 100 μg/mL streptomycin (Gibco), and 0.5 μg/mL puromycin (Sigma).

Techniques: Modification, Control

Journal: Microbiology Spectrum

Article Title: Epitranscriptomic N 6 -Methyladenosine Profile of SARS-CoV-2-Infected Human Lung Epithelial Cells

doi: 10.1128/spectrum.03943-22

Figure Lengend Snippet: Selected differentially m 6 A-modified transcripts in SARS-CoV-2-infected A549-hACE2 cells versus mock-infected control cells by m 6 A quantity ^a

Article Snippet: A549-hACE2 lung carcinoma cells expressing the human ACE2 protein (Invivogen) were maintained in DMEM with 4.5 g/L glucose and 2 mM l -glutamine (Gibco), 10% heat-inactivated fetal bovine serum (FBS; R&D Systems), 100 U/mL penicillin, 100 μg/mL streptomycin (Gibco), and 0.5 μg/mL puromycin (Sigma).

Techniques: Control

Journal: Microbiology Spectrum

Article Title: Epitranscriptomic N 6 -Methyladenosine Profile of SARS-CoV-2-Infected Human Lung Epithelial Cells

doi: 10.1128/spectrum.03943-22

Figure Lengend Snippet: Selected differentially m 6 A-modified transcripts in SARS-CoV-2-infected A549-hACE2 cells versus mock-infected control cells by percent modified ^a

Article Snippet: A549-hACE2 lung carcinoma cells expressing the human ACE2 protein (Invivogen) were maintained in DMEM with 4.5 g/L glucose and 2 mM l -glutamine (Gibco), 10% heat-inactivated fetal bovine serum (FBS; R&D Systems), 100 U/mL penicillin, 100 μg/mL streptomycin (Gibco), and 0.5 μg/mL puromycin (Sigma).

Techniques: Control, Modification

Journal: Microbiology Spectrum

Article Title: Epitranscriptomic N 6 -Methyladenosine Profile of SARS-CoV-2-Infected Human Lung Epithelial Cells

doi: 10.1128/spectrum.03943-22

Figure Lengend Snippet: Validation of differential methylation for selected transcripts in A549-hACE2 cells. (A) Schematic of m 6 A RIP. A549-hACE2 cells were mock infected or infected with ΔS-VRP(G) for 24 h. Total RNA was used for m6A-RIP. (B) Total RNA from cells infected with ΔS-VRP(G) was used for m 6 A-RIP. The fold enrichment of m 6 A-modfied RNA in the IP fraction of infected cell RNA versus mock-infected controls is indicated. (C) Total cellular RNA from mock-infected or ΔS-VRP(G)-infected cells was used for RT-qPCR analysis of total levels of mature miR-4486. (D) Total cellular RNA from mock-infected or ΔS-VRP(G)-infected cells was used for m 6 A RNA-IP to determine the levels of m 6 A-modified mature miR-4486. (C and D) Statistical significance was determined by t test compared to mock control. *, P < 0.05; **, P < 0.005; ***, P < 0.0005.

Article Snippet: A549-hACE2 lung carcinoma cells expressing the human ACE2 protein (Invivogen) were maintained in DMEM with 4.5 g/L glucose and 2 mM l -glutamine (Gibco), 10% heat-inactivated fetal bovine serum (FBS; R&D Systems), 100 U/mL penicillin, 100 μg/mL streptomycin (Gibco), and 0.5 μg/mL puromycin (Sigma).

Techniques: Methylation, Infection, Quantitative RT-PCR, Modification, Control

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: Pharmaceuticals

Article Title: Improved HDAC Inhibition, Stronger Cytotoxic Effect and Higher Selectivity against Leukemias and Lymphomas of Novel, Tricyclic Vorinostat Analogues

doi: 10.3390/ph14090851

Figure Lengend Snippet: IC 50 [µM] of Vorinostat derivatives 7a-t based on the survival of non-cancerous (BALB/3T3) and cancerous (MV4-11, Daudi, MCF-7 and A549) cells after 72 h of treatment. N/T—not tested.

Article Snippet: Human biphenotypic B myelomonocytic leukemia MV4-11 and normal mouse fibroblast BALB/3T3 cell line were obtained from American Type Culture Collection (USA); human lung carcinoma A549 cell line and human adenocarcinoma breast cancer MCF-7 cell line were obtained from European Collection of Authenticated Cell Cultures (UK).

Techniques:

Journal: Pharmaceuticals

Article Title: Improved HDAC Inhibition, Stronger Cytotoxic Effect and Higher Selectivity against Leukemias and Lymphomas of Novel, Tricyclic Vorinostat Analogues

doi: 10.3390/ph14090851

Figure Lengend Snippet: Selectivity index (IC 50 of normal vs. cancer cells). SI > 1.0 indicates a compound of greater activity against cancer cells and lower cytotoxicity on normal cells.

Article Snippet: Human biphenotypic B myelomonocytic leukemia MV4-11 and normal mouse fibroblast BALB/3T3 cell line were obtained from American Type Culture Collection (USA); human lung carcinoma A549 cell line and human adenocarcinoma breast cancer MCF-7 cell line were obtained from European Collection of Authenticated Cell Cultures (UK).

Techniques: Activity Assay

Journal: Pharmaceutics

Article Title: Effect of E. cava and C. indicum Complex Extract on Phorbol 12-Myristate 13-Acetate (PMA)-Stimulated Inflammatory Response in Human Pulmonary Epithelial Cells and Particulate Matter (PM) 2.5 -Induced Pulmonary Inflammation in Mice

doi: 10.3390/pharmaceutics15112621

Figure Lengend Snippet: Effects of cell viability and MUC5AC mRNA levels of ED and dieckol. ( A ) In a 96-well plate, 1 × 10 4 cells per well were seeded. After 24 h, ED and dieckol were treated for another 24 h with serum-free medium. Viability was measured using MTT assay. ( B ) RT-PCRanalysis using Rig/S15 as the loading control was performed for measurement of MUC5AC mRNA expression in A549 cells. After ED and dieckol pre-treatment for 30 min, cells were PMA-stimulated for 24 h. The relative mRNA levels of MUC5AC were quantified using the Image J program. The average value of three independent experiments is shown. All data are expressed as the mean ± SD of the experiment. ### p < 0.001 compared to the control group. ** p < 0.01 and *** p < 0.001, compared to the PMA control group. ns: not statistically significant.

Article Snippet: The A549 pulmonary epithelial cell line was obtained from the Korean Cell Line Bank (KCLB; Seoul, Republic of Korea).

Techniques: MTT Assay, Control, Expressing

Journal: Pharmaceutics

Article Title: Effect of E. cava and C. indicum Complex Extract on Phorbol 12-Myristate 13-Acetate (PMA)-Stimulated Inflammatory Response in Human Pulmonary Epithelial Cells and Particulate Matter (PM) 2.5 -Induced Pulmonary Inflammation in Mice

doi: 10.3390/pharmaceutics15112621

Figure Lengend Snippet: Effect of ED and dieckol on the phosphorylation of MAPKs in PMA-stimulated A549 cells. ( A ) ED and ( B ) dieckol pre-treatment for 30 min; cells were PMA-stimulated for 30 min. β-actin were detected and used as internal controls. The relative protein levels of p-JNK, p-ERK, and p-p38 were quantified using the Image J program and normalized to β-actin. The average value of three independent experiments is shown. All data are expressed as the mean ± SD of the experiment. ## p < 0.01 and ### p < 0.001 compared to the control group; * p < 0.05, ** p < 0.01, and *** p < 0.001 compared to the PMA control group. See also .

Article Snippet: The A549 pulmonary epithelial cell line was obtained from the Korean Cell Line Bank (KCLB; Seoul, Republic of Korea).

Techniques: Phospho-proteomics, Control