a549 cells  (Genecopoeia)

Journal: OncoTargets and therapy

Article Title: MiR-21-5p promotes the progression of non-small-cell lung cancer by regulating the expression of SMAD7

doi: 10.2147/OTT.S172393

Figure Lengend Snippet: MiR-21-5p promoted the invasion/migration of NSCLC cells. Notes: ( A ) A higher expression of MiR-21-5p was associated with a stronger migration ability of A549 cells. ( B ) A higher expression of MiR-21-5p was associated with a stronger invasion ability of A549 cells. ( C ) Detection of EMT-related protein expressions (MMP-9, E-cadherin, and vimentin). * P

Article Snippet: The expression level of MiR-21-5p in the human NSCLC cell line A549 transfected with or without MiR-21- 5p-inhibitor/mimic RNA (MiR-21-5p-NC) was detected by the miRNA-qPCR kit (GeneCopoeia Inc, Germantown, MD, USA) and a real-time fluorescence quantitative PCR instrument (Agilent Technologies).

Techniques: Migration, Expressing

Journal: Free radical biology & medicine

Article Title: Enhanced Mitochondrial DNA Repair of the Common Disease- Associated Variant, Ser326Cys, of hOGG1 through Small Molecule Intervention

doi: 10.1016/j.freeradbiomed.2018.05.094

Figure Lengend Snippet: OGG1 activators attenuate paraquat-induced oxidative damage in A549 cells A549 cells were pre-treated for 4 hr with small molecule OGG1 activators or 0.1% DMSO prior to addition of 0.6 mM paraquat for 48 hr. The cells were stained and imaged for 8-oxoG content and the average fluorescence intensity within the cell was normalized based on the number of cells imaged. (A) Graphical representations of the change in 8-oxoG intensity within the cytoplasm of cells compared to 0.6 mM paraquat treated cells (dashed line). (B) Representative images of cells from panel A depict changes in mitochondrial 8-oxoG staining (Hoechst 33342, overlaid blue; MitoTracker® orange, overlaid orange; 8-oxoG-AleaFluor®-647, overlaid red; line = 40 μm). (C) The percent of the cell populations labeled as high responders is shown for A549 cells pre-incubated with OGG1 activators (4 hr) prior to exposure to a single concentration of paraquat (0.6 mM, dashed line) for 48 hr. (D) The images from panel A were analyzed further by examining morphology changes. The mean nuclear area (dashed line indicates no treatment, 0.6 mM PQ) for A549 cells was graphed depicting a change in overall cell health. Significance was determined using a 1-way ANOVA with a Dunnett’s posttest comparing paraquat alone treated groups to untreated control (0 mM) and OGG1 activator treated groups to 0.6 mM paraquat control (** p

Article Snippet: To alter the content of OGG1 in A549 cells, BacMam virus constructs were generated using expression vector, pDONRhumanOGG1v1a_FL (GeneCopoeia; Rockville, MD).

Techniques: Staining, Fluorescence, Labeling, Incubation, Concentration Assay

Journal: Free radical biology & medicine

Article Title: Enhanced Mitochondrial DNA Repair of the Common Disease- Associated Variant, Ser326Cys, of hOGG1 through Small Molecule Intervention

doi: 10.1016/j.freeradbiomed.2018.05.094

Figure Lengend Snippet: OGG1 activators protect against paraquat-induced loss of mitochondrial membrane potential in A549 cells A549 cells were pre-treated for 4 hours with the OGG1 small molecule activators or 0.1% DMSO prior to exposure to 0.3 mM paraquat for 24 hr. Mitochondrial membrane potential was measured from images captured as described in the methods. (A) Graphical representation of the change in the JC-1 ratio with increasing concentration of paraquat and cells pre-treated with the OGG1 activators prior to exposure to a single concentration of paraquat (0.3 mM) for 24 hr. (B) Representative images from untreated, 3 mM paraquat, 0.3 mM paraquat, and Compound D (30 μM) with 0.3 mM paraquat exposed cells (Hoechst 33342, overlaid blue; JC-1 monomer, overlaid green; JC-1 aggregate, overlaid orange; CellMask™ Deep Red, overlaid red; line = 40 μm). Significance was determined using a 1-way ANOVA with a Dunnett’s posttest comparing paraquat alone treated groups to untreated control (0 mM) and OGG1 activator treated groups to 0.3 mM paraquat control (* p

Article Snippet: To alter the content of OGG1 in A549 cells, BacMam virus constructs were generated using expression vector, pDONRhumanOGG1v1a_FL (GeneCopoeia; Rockville, MD).

Techniques: Concentration Assay

Journal: Free radical biology & medicine

Article Title: Enhanced Mitochondrial DNA Repair of the Common Disease- Associated Variant, Ser326Cys, of hOGG1 through Small Molecule Intervention

doi: 10.1016/j.freeradbiomed.2018.05.094

Figure Lengend Snippet: Paraquat-induced mitochondrial DNA damage is inversely related to OGG1 expression A549 cells were treated with paraquat for 24 or 48 hr and subjected to formaldehyde fixation prior to immunofluorescence detection of 8-oxoguanine. (A) Graphical representation of the percent change in 8-oxoguanine intensity within the cytoplasm region following 24- and 48-hr treated cells with increasing concentrations of paraquat. Significance was determined using a 1-way ANOVA with a Dunnett’s posttest comparing treatment groups to an untreated control (* p

Article Snippet: To alter the content of OGG1 in A549 cells, BacMam virus constructs were generated using expression vector, pDONRhumanOGG1v1a_FL (GeneCopoeia; Rockville, MD).

Techniques: Expressing, Immunofluorescence

Journal: Molecular cell

Article Title: Combinatorial CRISPR-Cas9 metabolic screens reveal critical redox control points dependent on the KEAP1-NRF2 regulatory axis

doi: 10.1016/j.molcel.2018.01.017

Figure Lengend Snippet: Combinatorial CRISPR screens reveal metabolic network dependencies (A) SKO fitness scores for HeLa cells, plotted as f g (day −1 ), with a more negative score representing a loss in fitness with SKO. Plotted as mean ± SD. (B) Multi-isoform family member fitness scores and gene expression for HeLa (top) and A549 (bottom) cells. (C) Relative comparison of SKO fitness scores (f g ) across both cells. (D) Relative comparison of genetic interaction scores (π gg .

Article Snippet: CRISPR Cas9 nuclease stable expressing HeLa and A549 cells were obtained from GeneCopoeia and grown in DMEM medium with 10% FBS and Antibiotic-Antimycotic.

Techniques: CRISPR, Expressing

Journal: Oncogene

Article Title: MicroRNA-24 regulates XIAP to reduce the apoptosis threshold in cancer cells

doi: 10.1038/onc.2012.258

Figure Lengend Snippet: miR-24 modulates XIAP expression. ( A ) Transfection of pre-miR-24 in A549 cells results in a 23-fold increase in miR-24 expression compared to miR negative control. Bars represent miR-24 expression relative to control and standard deviation from triplicate experiments (**p

Article Snippet: To establish A549/XIAP-GFP clones, A549 cells were transduced with lentiviral p-Receiver-lv19-XIAP (XIAP-GFP) or p-Receiver-lv19 empty vector (GFP-empty vector) (GeneCopoeia, Rockville, MD).

Techniques: Expressing, Transfection, Negative Control, Standard Deviation

Journal: Oncogene

Article Title: MicroRNA-24 regulates XIAP to reduce the apoptosis threshold in cancer cells

doi: 10.1038/onc.2012.258

Figure Lengend Snippet: Chromosomal loss in the miR-24 coding region of A549 and H1437. ( A ) Summary of array-CGH data from chromosome 19 in H1437, A549, and CALU-1 NSCLC cells. The miR-24 coding region is located at 19p13.13. Genomic loss is represented by bars to the left of center or negative Log2 ratios while genomic gain is represented by bars to the right of center or positive Log2 ratios. ( B ) Representative spectral karyotyping (SKY) analysis of chromosome 19 in A549 (upper) and H1437 (lower) lung cancer cell lines. A549 cells carry a partial p-arm deletion in chromosome 19. H1437 has monosomy of chromosome 19 in over 40% of cells analyzed.

Article Snippet: To establish A549/XIAP-GFP clones, A549 cells were transduced with lentiviral p-Receiver-lv19-XIAP (XIAP-GFP) or p-Receiver-lv19 empty vector (GFP-empty vector) (GeneCopoeia, Rockville, MD).

Techniques:

Journal: Oncogene

Article Title: MicroRNA-24 regulates XIAP to reduce the apoptosis threshold in cancer cells

doi: 10.1038/onc.2012.258

Figure Lengend Snippet: Over-expression of miR-24 targets endogenous but not ectopic XIAP in A549/XIAP-GFP stably transfected cells. ( A, B ) Stable cell line A549/GFP-empty vector or A549/XIAP-GFP without the 3′ UTR of XIAP was transfected with pre-miR-24 or miR negative control for 48 hours, and then incubated with 100 ng/ml of TRAIL for additional 48 hours. ( A ) Representative western blot of endogenous and ectopic XIAP protein levels. miR-24 over-expression reduces endogenous XIAP protein levels but has no effect on ectopic XIAP protein levels. ( B ) MTT cell viability assay. Ectopic expression of the XIAP-GFP fusion protein inhibits the effect of pre-miR-24 and TRAIL treatment on cell viability. Bars represent cell viability as a percentage relative to untreated cells and standard deviation from triplicate experiments (**p

Article Snippet: To establish A549/XIAP-GFP clones, A549 cells were transduced with lentiviral p-Receiver-lv19-XIAP (XIAP-GFP) or p-Receiver-lv19 empty vector (GFP-empty vector) (GeneCopoeia, Rockville, MD).

Techniques: Over Expression, Stable Transfection, Transfection, Plasmid Preparation, Negative Control, Incubation, Western Blot, MTT Assay, Viability Assay, Expressing, Standard Deviation

Journal: Oncogene

Article Title: MicroRNA-24 regulates XIAP to reduce the apoptosis threshold in cancer cells

doi: 10.1038/onc.2012.258

Figure Lengend Snippet: miR-24-mediated XIAP suppression restores TRAIL-induced apoptosis in TRAIL-resistant cells. ( A ) Cell viability of A549 ( i ), H292 ( ii ) OE21 ( iii ), HeLa ( iv ), SK-BR-3 ( v ), and MDA-MB-468 ( vi ) cells following a 48-hour transfection with pre-miR-24 or miR-negative control and an additional 48h treatment with increasing concentrations of soluble TRAIL (*p

Article Snippet: To establish A549/XIAP-GFP clones, A549 cells were transduced with lentiviral p-Receiver-lv19-XIAP (XIAP-GFP) or p-Receiver-lv19 empty vector (GFP-empty vector) (GeneCopoeia, Rockville, MD).

Techniques: Multiple Displacement Amplification, Transfection, Negative Control

Journal: Oncogene

Article Title: MicroRNA-24 regulates XIAP to reduce the apoptosis threshold in cancer cells

doi: 10.1038/onc.2012.258

Figure Lengend Snippet: Down-regulation of mature miR-24 correlates with increased XIAP protein levels. ( A ) Real-time RT-PCR analysis of mature miR-24 expression in normal (SAEC), NSCLC (CALU-1, A549, H1437, H292), cervical carcinoma (HeLa), esophageal (OE21) and breast (SK-BR-3, MDA-MB-468) cancer cells. Bars represent mean and standard deviation from triplicate experiments. ( B ) Representative western blot of XIAP protein in normal and cancer cells. Quantitative XIAP protein levels are normalized to SAEC. GAPDH served as a loading control. ( C–E ) Real-time RT-PCR quantification of the primary (Pri-miR) miR-24-2 cluster located on chromosome 19 (C), which also contains mature miR-23a (D) and mature miR-27a (E). Bars represent mean and standard deviation from triplicate experiments.

Article Snippet: To establish A549/XIAP-GFP clones, A549 cells were transduced with lentiviral p-Receiver-lv19-XIAP (XIAP-GFP) or p-Receiver-lv19 empty vector (GFP-empty vector) (GeneCopoeia, Rockville, MD).

Techniques: Quantitative RT-PCR, Expressing, Multiple Displacement Amplification, Standard Deviation, Western Blot