Journal: International Journal of Molecular Sciences

Article Title: Targeted Phagocytosis Induction for Cancer Immunotherapy via Bispecific MerTK-Engaging Antibodies

doi: 10.3390/ijms232415673

Figure Lengend Snippet: Characterisation of parental MerTK antibodies. Kinetics determination against soluble hMerTK-TS for either ( a ) MerK28 or ( b ) oaMerK28, and against mMerTK-Fc for ( c ) MerK28 or ( d ) oaMerK28. ( e , f ) Activation of signalling transduction with MerK28. pAKT HTRF assays with either A431 ( e ) or HEK293 ( f ) cells. Unstimulated cells are shown as “NC”, and the bivalent MerK28 and one-armed MerK28 (oaMerK28) are plotted. Error bars represent biological duplicate measurements. ( g ) Epitope binning of MerK28 (blue) and MerK17 (green) via BLI. Human MerTK-His 6 was immobilised on HIS1K biosensors and associated to the indicated antibodies for 600 s each. PBS is shown in black. ( h ) Ligand competition assay for MerK28. Association to hMerTK (light blue) alone or to MerTK pre-incubated with Pros1 ligand (pink).

Article Snippet: The following cell lines were purchased from DSMZ (Brunswick, Germany): human epidermoid carcinoma A431 (ACC 91), human embryonic kidney HEK293 (ACC 305), human lung carcinoma A549 (ACC 107), and human acute monocytic leukaemia THP-1 (ACC 16).

Techniques: Activation Assay, Transduction, Competitive Binding Assay, Incubation

Journal: International Journal of Molecular Sciences

Article Title: Targeted Phagocytosis Induction for Cancer Immunotherapy via Bispecific MerTK-Engaging Antibodies

doi: 10.3390/ijms232415673

Figure Lengend Snippet: Bispecific MerTK/EGFR antibodies and their binding capabilities. ( a ) Simultaneous binding of hMerTK and EGFR via BLI. Antibodies were loaded onto AHC biosensors and then associated first to MerTK and then to EGFR for 300 s. The colour-coding is the same as represented in the legend for ( d ). Cell titration on ( b , c ) A431 and ( d ) HEK293 cell lines by staining the antibodies with anti-human Fc PE and plotting the relative fluorescence units (RFU) versus the antibody concentration. The negative controls incubated only with secondary antibodies are shown as “NC”. Error bars represent biological duplicates.

Article Snippet: The following cell lines were purchased from DSMZ (Brunswick, Germany): human epidermoid carcinoma A431 (ACC 91), human embryonic kidney HEK293 (ACC 305), human lung carcinoma A549 (ACC 107), and human acute monocytic leukaemia THP-1 (ACC 16).

Techniques: Binding Assay, Titration, Staining, Fluorescence, Concentration Assay, Incubation

Journal: International Journal of Molecular Sciences

Article Title: Targeted Phagocytosis Induction for Cancer Immunotherapy via Bispecific MerTK-Engaging Antibodies

doi: 10.3390/ijms232415673

Figure Lengend Snippet: Macrophage-engagement and phagocytosis of tumour cells. ( a ) Contour plots of A431 binding on the x-axis (FITC fluorescence) and THP-1 binding on the y-axis (PE fluorescence) following the colour-coding of ( b ). Double-positive events in the upper-right Q2 quadrant show simultaneous engagement of THP-1 and A431 cells, whereas the upper-left Q1 quadrant represents MerTK-binding only. ( b ) Frequency of parent of Q1 and Q2 from ( a ). Coloured squares represent incubation of antibodies only on THP-1 cells, whereas the triangles represent co-incubation of pre-stained THP-1 cells and Calcein-AM-stained A431 cells. ( c ) Quantitation of CD14+-THP-1 cells and Calcein-AM red—orange stained A431 cells for phagocytosis assays. The negative control (NC) represents co-incubation of both cell types without antibody treatment. ( d ) Determination of phagocytosis rate (%) after co-incubation of THP-1 cells in different differentiation states together with A431 cells. Error bars represent the standard deviation of at least biological duplicates.

Article Snippet: The following cell lines were purchased from DSMZ (Brunswick, Germany): human epidermoid carcinoma A431 (ACC 91), human embryonic kidney HEK293 (ACC 305), human lung carcinoma A549 (ACC 107), and human acute monocytic leukaemia THP-1 (ACC 16).

Techniques: Binding Assay, Fluorescence, Incubation, Staining, Quantitation Assay, Negative Control, Standard Deviation

Journal: Radiation Oncology (London, England)

Article Title: The use of caspase inhibitors in pulsed-field gel electrophoresis may improve the estimation of radiation-induced DNA repair and apoptosis

doi: 10.1186/1748-717X-6-6

Figure Lengend Snippet: PFGE pattern . (A) PFGE for a typical A431 cell line experiment with agarose-encapsulated cells. The range of DNA quantification, beginning just before the compression zone (CZ) and ending at the bottom of the smears, is shown. (B) The amount of released DNA (optical density, Arbitrary Units, AU) was plotted as a function of time using the following time-points 0, 0.5, 1, 2 or 4 h. U stands for unirradiated (black bars) and I for 45 Gy irradiated cell-plugs (white bars). Data were obtained from one experiment.

Article Snippet: The tumour cells used in this study were human squamous carcinoma A431 from the American Type Cell Collection (LGC Promochem, Barcelona, Spain), and human pancreatic carcinoma NP18 cell line.

Techniques: Irradiation

Journal: Radiation Oncology (London, England)

Article Title: The use of caspase inhibitors in pulsed-field gel electrophoresis may improve the estimation of radiation-induced DNA repair and apoptosis

doi: 10.1186/1748-717X-6-6

Figure Lengend Snippet: Caspase inhibitors blocked apoptosis induced by agarose encapsulation . Representative PFGE from A431 and NP18 cell lines are depicted. Spontaneous DNA fragmentation induced exclusively by incubation in unirradiated cell-plugs (black bars) was inhibited (grey bars) by zvad-fmk (100 μM). * P = 0.016; ** P = 0.083 (Mann-Whitney test, 4 independent experiments). No significant differences were found between the A431 and N18 cell lines.

Article Snippet: The tumour cells used in this study were human squamous carcinoma A431 from the American Type Cell Collection (LGC Promochem, Barcelona, Spain), and human pancreatic carcinoma NP18 cell line.

Techniques: Encapsulation, Incubation, MANN-WHITNEY

Journal: BioMed Research International

Article Title: Photosensitizer Adhered to Cell Culture Microplates Induces Phototoxicity in Carcinoma Cells

doi: 10.1155/2013/549498

Figure Lengend Snippet: Experimental schemes. (a) The distribution of PS from microplates that were preincubated with PSs and then incubated with A431 cells was analyzed in three microplates (MP 1, MP 2, and MP 3). (b) Different incubation protocols were applied prior to photodynamic treatment to assess the phototoxicity of microplate-adhered PS. Abbreviations: DPBS: Dulbecco's phosphate-buffered saline; FBS: fetal bovine serum; MP: microplate; PS: photosensitizer; SN: supernatant.

Article Snippet: A431 human epidermoid carcinoma cells (ATCC-Nr. CRL-1555) were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10 mM HEPES, 5% (v/v) fetal bovine serum (FBS), 2 mM L-glutamine, 1 mM Na-pyruvate, 100 U mL −1 penicillin, and 0.1 mg mL −1 streptomycin.

Techniques: Incubation, Saline