Journal: PLoS ONE

Article Title: Bacterial Delivery of TALEN Proteins for Human Genome Editing

doi: 10.1371/journal.pone.0091547

Figure Lengend Snippet: ( A ) HeLa-Venus cells were infected with the indicated strains at an MOI of 100 for 3 hours. Nuclear proteins were extracted and subjected to SDS PAGE and Western blot by anti-Flag antibody; ( B ) HeLa-Venus cells were infected with PAK-JΔ STY /pExoS54-F-TALEN1 and PAK-JΔ STY /pExoS54-F-TALEN2 at MOI of 100 each for 3 hours. At the indicated time after the termination of infection, cytoplasmic and nuclear protein extracts were prepared and subjected to SDS PAGE and Western blot assay using anti-Flag Ab. NI, no infection control.

Article Snippet: Human cervix epithelial cell line HeLa (ATCC CCL-2) was grown in Dulbecco’s Modified Eagle Medium (DMEM; Gibco) supplemented with 10% heat-inactivated Fetal Bovine Serum (FBS).

Techniques: Infection, SDS Page, Western Blot, Control

Journal: PLoS ONE

Article Title: Bacterial Delivery of TALEN Proteins for Human Genome Editing

doi: 10.1371/journal.pone.0091547

Figure Lengend Snippet: ( A ) Fluorescent intensity of HeLa-Venus cells transfected by the indicated TALEN encoding plasmid constructs. Cells were analyzed by flow cytometry 3 days after the transfection. ( B ) Sequence changes in TALEN target region among the Venus negative cells.

Article Snippet: Human cervix epithelial cell line HeLa (ATCC CCL-2) was grown in Dulbecco’s Modified Eagle Medium (DMEM; Gibco) supplemented with 10% heat-inactivated Fetal Bovine Serum (FBS).

Techniques: Transfection, Plasmid Preparation, Construct, Flow Cytometry, Sequencing

Journal: PLoS ONE

Article Title: Bacterial Delivery of TALEN Proteins for Human Genome Editing

doi: 10.1371/journal.pone.0091547

Figure Lengend Snippet: ( A ) Multiple rounds of TALEN injection. HeLa-Venus cells were infected with PAK-JΔ STY /pExoS54-F-TALEN1 and PAK-JΔ STY /pExoS54-F-TALEN2 at MOI of 100 each for 3 hours. Bacteria were cleared and cells were cultured for 24 hours, then repeated injection at MOI of 50 or 100. The FACS data of infected cells were compared by paired t-test and ANOVA. *P<0.05; **P<0.001. Error bars indicate standard deviations of triplicate assays. ( B ) HeLa-Venus cells were either left unsynchronized (U) or synchronized (S) through double thymidine blocking and released to determine the duration of the cell cycle by FACS analysis. ( C ) HeLa-Venus cells were synchronized to S phase (S) or unsynchronized (U) and infected by PAK-JΔ STY /pExoS54-F-TALEN1 and PAK-JΔ STY /pExoS54-F-TALEN2 at MOI of 100 each for 3 hours. The FACS data of infected cell populations were compared by paired or two-sample t-test. *P<0.05; **P<0.001. Error bars indicated the standard deviations of triplicate assays.

Article Snippet: Human cervix epithelial cell line HeLa (ATCC CCL-2) was grown in Dulbecco’s Modified Eagle Medium (DMEM; Gibco) supplemented with 10% heat-inactivated Fetal Bovine Serum (FBS).

Techniques: Injection, Infection, Bacteria, Cell Culture, Blocking Assay

Journal: PLoS ONE

Article Title: Amiloride-enhanced gene transfection of octa-arginine functionalized calcium phosphate nanoparticles

doi: 10.1371/journal.pone.0188347

Figure Lengend Snippet: a) Schematic concept of fabrication of the octa-arginine-functionalized calcium phosphate nanoparticles. b) Scanning electron micrograph (SEM) and transmission electron micrograph (TEM) of CaP nanoparticles functionalized with octa-arginine (CaP/DNA/CaP/R8). c) Transmission light microscopy (TLM) and fluorescence microscopy (FM) of transfected cells. Representative images of HeLa, Saos-2, hMSC, and hOB cells transfected with the pAcGFP1 plasmid within CaP nanoparticles functionalized with octa-arginine at a concentration of 50 mg ml −1 . Transfected cells appear green under fluorescence microscopy. Magnification: x20. Bar = 100 μm.

Article Snippet: For the transfection experiments, human transformed cervix epithelial cells (HeLa) (TaKaRa Bio Co.), primary osteosarcoma (Saos-2) cells (TaKaRa), hMSCs (Cellular Engineering Technologies), and hOBs derived from hipbone (PromoCell GmbH) were cultured in DMEM, supplemented with 10% foetal bovine serum (HyClone ® , Thermo Scientific), and 1% Pen Strep (10,000 U ml −1 penicillin / 10,000 μg ml −1 streptomycin) (Gibco; Life Technologies) at 37°C in a humidified atmosphere with 5% CO 2 .

Techniques: Transmission Assay, Light Microscopy, Fluorescence, Microscopy, Transfection, Plasmid Preparation, Concentration Assay

Journal: PLoS ONE

Article Title: Amiloride-enhanced gene transfection of octa-arginine functionalized calcium phosphate nanoparticles

doi: 10.1371/journal.pone.0188347

Figure Lengend Snippet: Transfection efficiency (TE) and cell viability (CV) in HeLa, Saos-2, hMSC, and hOB cells.

Article Snippet: For the transfection experiments, human transformed cervix epithelial cells (HeLa) (TaKaRa Bio Co.), primary osteosarcoma (Saos-2) cells (TaKaRa), hMSCs (Cellular Engineering Technologies), and hOBs derived from hipbone (PromoCell GmbH) were cultured in DMEM, supplemented with 10% foetal bovine serum (HyClone ® , Thermo Scientific), and 1% Pen Strep (10,000 U ml −1 penicillin / 10,000 μg ml −1 streptomycin) (Gibco; Life Technologies) at 37°C in a humidified atmosphere with 5% CO 2 .

Techniques: Transfection

Journal: PLoS ONE

Article Title: Amiloride-enhanced gene transfection of octa-arginine functionalized calcium phosphate nanoparticles

doi: 10.1371/journal.pone.0188347

Figure Lengend Snippet: A) HeLa, b) Saos-2, c) hMSC, and d) hOB cells transfected with CaP nanoparticles loaded with pUC57 plasmid and functionalized with different concentrated solutions of R8 (0.1, 1, 5, 10, 50, 100 mg ml −1 ), PEI, and protamine. pUC57 group are single shell non-functionalized CaP nanoparticles. The bars represent the mean ± standard deviation. *p < 0.05 compared to transfection of non-pre-treated cells (NT).

Article Snippet: For the transfection experiments, human transformed cervix epithelial cells (HeLa) (TaKaRa Bio Co.), primary osteosarcoma (Saos-2) cells (TaKaRa), hMSCs (Cellular Engineering Technologies), and hOBs derived from hipbone (PromoCell GmbH) were cultured in DMEM, supplemented with 10% foetal bovine serum (HyClone ® , Thermo Scientific), and 1% Pen Strep (10,000 U ml −1 penicillin / 10,000 μg ml −1 streptomycin) (Gibco; Life Technologies) at 37°C in a humidified atmosphere with 5% CO 2 .

Techniques: Transfection, Plasmid Preparation, Standard Deviation

Journal: Oncotarget

Article Title: Inhibition of succinate dehydrogenase by the mitochondrial chaperone TRAP1 has anti-oxidant and anti-apoptotic effects on tumor cells

doi:

Figure Lengend Snippet: (A-C) TRAP1 expression levels were measured in SAOS-2 osteosarcoma cells (A), HeLa cervix carcinoma cells (B) and mouse embryo fibroblasts (MEF; C) by Western immunoblot assays. GAPDH was used as a loading control. SAOS-2 and Hela cells were transfected either with a scrambled shRNA (mock) or with two different TRAP1 shRNAs (shTRAP1a and shTRAP1b); shTRAP1b cells transfected with a mouse TRAP1 cDNA insensitive to interference by shTRAP1 were indicated as shTRAP1b + mTRAP1. MEFs were stably transfected with either a TRAP1 cDNA or with an empty vector (mock). (D-F) Intracellular hydrogen peroxide levels were assessed by Amplex Red in lysates from SAOS-2 (D), HeLa (E) or MEF (F) cells. (G-I) Cytofluorimetric analysis of mitochondrial superoxide levels was carried out by staining SAOS-2 (G), HeLa (H) or MEF (I) cells with the fluorescent probe MitoSox. All data are reported as mean±SD values (n≥3; *P<0.05 in a Student's t test).

Article Snippet: Human SAOS-2 osteosarcoma cells and human HeLa cervix carcinoma cells were purchased from ATCC.

Techniques: Expressing, Western Blot, Control, Transfection, shRNA, Stable Transfection, Plasmid Preparation, Staining

Journal: PLoS ONE

Article Title: Lymphocyte Display: A Novel Antibody Selection Platform Based on T Cell Activation

doi: 10.1371/journal.pone.0007174

Figure Lengend Snippet: (A) Cell-surface expression of CEACAM5 (CEA) and NIP-modified molecules on HeLa, HeLa CEA and HeLa cells labeled with 2.5 µg/mL of the hapten (HeLa NIP ). (B) FACS analysis of CD69 expression by Jurkat EGFP , Jurkatα CEA-CIR-EGFP and Jurkatα NIP-CIR stimulated either with immobilized anti-CD3 mAb or target cells (E∶T = 1∶1; HeLa, HeLa CEA or HeLa NIP ) for 16 hours.

Article Snippet: 293T cells (human embryo kidney epithelia; CRL-11268), HeLa cells (human cervix carcinoma; CCL-2), HT-1080 cells (human fibrosarcoma; CCL-121), MKN45 cells (human stomach adenocarcinoma; JCRB-0254), and MDA-MB231 cells (human breast adenocarcinoma; HTB-26) were grown in DMEM supplemented with 10% (vol/vol) heat-inactivated FCS, (Invitrogen Life Technologies, Gaithersburg, MD, USA), referred as to DMEM complete medium (DCM).

Techniques: Expressing, Modification, Labeling

Journal: PLoS ONE

Article Title: Lymphocyte Display: A Novel Antibody Selection Platform Based on T Cell Activation

doi: 10.1371/journal.pone.0007174

Figure Lengend Snippet: (A) Cell-surface expression of CEACAM5 (CEA) on HeLa, HT1080, MDA-MB-231 and MKN45 cells. (B) FACS analysis of CD69 expression by Jurkat, Jurkatα CEA-CIR-EGFP and Jurkatα NIP-CIR stimulated either with immobilized anti-CD3 mAb or target cells (E∶T = 1∶1; HeLa, HT1080, MDA-MB-231 or MKN45) for 16 hours.

Article Snippet: 293T cells (human embryo kidney epithelia; CRL-11268), HeLa cells (human cervix carcinoma; CCL-2), HT-1080 cells (human fibrosarcoma; CCL-121), MKN45 cells (human stomach adenocarcinoma; JCRB-0254), and MDA-MB231 cells (human breast adenocarcinoma; HTB-26) were grown in DMEM supplemented with 10% (vol/vol) heat-inactivated FCS, (Invitrogen Life Technologies, Gaithersburg, MD, USA), referred as to DMEM complete medium (DCM).

Techniques: Expressing

Journal: Journal of Biological Inorganic Chemistry

Article Title: Revisiting the anticancer properties of phosphane(9-ribosylpurine-6-thiolato)gold(I) complexes and their 9H-purine precursors

doi: 10.1007/s00775-022-01968-x

Figure Lengend Snippet: Inhibitory concentrations IC 50 [µM] of 6-MP, 6-TG, 6-MMR, auranofin and complexes 1 – 9 when applied to EA.hy926 endothelial hybrid cells and cells of human HCT-116, HCT-116 p53−/− (p53 knockout mutant) and HT-29 colon carcinomas, HeLa and mdr KB-V1 Vbl cervix carcinoma (treated with and without 1 µM verapamil), MCF-7 and mdr MCF-7 Topo mamma carcinoma, U-87 glioblastoma, 518A2 melanoma, and human adult dermal fibroblasts HDFa

Article Snippet: 518A2 human melanoma cells (Department of Radiotherapy & Radiobiology, University Hospital Vienna, Austria), HCT116 wt (DSMZ ACC-581) and its HCT116 p53−/− knockout mutant colon carcinoma cells, U87 glioblastoma cells (ATCC HTB-14), EA.hy926 somatic cell hybrid cells (ATCC CRL-2922), HeLa cervix carcinoma cells (DSMZ ACC-57), MCF-7 breast cancer cells (DSMZ ACC-115), HT-29 cisplatin resistant colon cancer cells (DSMZ ACC-299) and non-malignant human dermal fibroblasts HDFa (ATCC PCS-201-012) were cultured in Dulbecco’s modified Eagle medium (PAN biotech), supplemented with 10% (v/v) fetal bovine serum (Sigma-Aldrich) and 1% (v/v) ZellShield (Minerva Biolabs) at 37 °C under 95% humidity and 5% CO 2 .

Techniques: Knock-Out, Mutagenesis

Journal: Cancers

Article Title: The Micro-Immunotherapy Medicine 2LPAPI ® Displays Immune-Modulatory Effects in a Model of Human Papillomavirus Type-16 L1-Protein Capsid-Treated Human Peripheral Blood Mononuclear Cells and Antiproliferative Effects in a Model of Cervical Cancer Cells

doi: 10.3390/cancers16071421

Figure Lengend Snippet: MIM-1 and MIM-5 lowered the cellular viability/proliferative potential in a model of starved HPV-positive cervical cancer cells. Cell viability/proliferative potential of HeLa cells was assessed via AlamarBlue reagent under the experimental conditions presented in (see ) when treated with the vehicle (Veh.), MIM-1 or MIM-5. The results are presented as the mean percentages ± standard deviation (S.D.) of n = 6 replicates per condition, with the Veh. being set at 100%.

Article Snippet: HeLa cells (human cervix epithelioid carcinoma) were supplied by the European Collection of Authenticated Cell Cultures (ECACC) (ref: #93021013, ECACC, Salisbury, UK) and cultured according to the supplier’s recommendations.

Techniques: Standard Deviation