Journal: FEBS letters

Article Title: Cyclin B1 transcription is enhanced by the p300 coactivator and regulated during the cell cycle by a CHR-dependent repression mechanism.

doi: 10.1016/s0014-5793(03)00028-0

Figure Lengend Snippet: Fig. 1. Cell cycle-dependent expression of chromosomally derived cyclin B1. NIH3T3 mouse ¢broblasts and HFF were serum-starved and re- stimulated by addition of serum to the medium. After 48 h of starvation, the 0 h time point represents cells mostly resting in G0. The following time points give results after serum restimulation. A: Cyclin B1 mRNA expression from NIH3T3 cells as measured by RT-PCR standardised relative to total RNA; averages of two experiments are given. B: FACS analyses of NIH3T3 cell populations at selected time points before and after serum addition. Cells were stained with propidium iodide and measured for DNA staining per cell. C: Expression of cyclin B1 mRNA in HFF cells with total RNA as standard quanti¢ed by RT-PCR. Averages of two experiments are given. D: Cyclin B1 protein expres- sion detected by immunoblotting. L-Actin served as a loading control.

Article Snippet: NIH3T3 cells (DSMZ, Braunschweig, Germany) for cell cycle analyses were cultured and transfected as previously described [19].

Techniques: Expressing, Derivative Assay, Reverse Transcription Polymerase Chain Reaction, Staining, Western Blot, Control

Journal: FEBS letters

Article Title: Cyclin B1 transcription is enhanced by the p300 coactivator and regulated during the cell cycle by a CHR-dependent repression mechanism.

doi: 10.1016/s0014-5793(03)00028-0

Figure Lengend Snippet: Fig. 3. Expression from a cyclin B1-promoter luciferase reporter comparing activity in G2/M versus resting cells. Mutation of the CHR leads to a loss of cell cycle regulation. Luciferase reporter plasmids carrying the human cyclin B1 upstream region together with a Renilla luciferase-expressing control plasmid were transfected into NIH3T3 ¢broblasts. Cells were synchronised by serum starva- tion for 66 h and restimulated. Ratios of luciferase activities from cell lysates 24 h after restimulation to activities in resting cells are given. Averages with standard deviations from six experiments are shown. Renilla luciferase was employed to standardise cyclin B1 ¢re- £y luciferase reporter expression.

Article Snippet: NIH3T3 cells (DSMZ, Braunschweig, Germany) for cell cycle analyses were cultured and transfected as previously described [19].

Techniques: Expressing, Luciferase, Activity Assay, Mutagenesis, Control, Plasmid Preparation, Transfection

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: ACS nano

Article Title: Colloidal Transformations in MS2 Virus Particles: Driven by pH, Influenced by Natural Organic Matter.

doi: 10.1021/acsnano.9b08112

Figure Lengend Snippet: Figure 3: A: Cryo-TEM image of MS2 at pH = 5.0 in water, aggregates are highlighted within red

Article Snippet: Virus propagation Page 17 of 30 ACS Paragon Plus Environment ACS Nano 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 18 MS2 (DSMZ 1367, DSMZ, Germany) was replicated in Escherichia coli strain W1485 (DSMZ 5695, DSMZ, Germany).

Techniques:

Journal: ACS nano

Article Title: Colloidal Transformations in MS2 Virus Particles: Driven by pH, Influenced by Natural Organic Matter.

doi: 10.1021/acsnano.9b08112

Figure Lengend Snippet: Figure 5: A: Cryo-TEM image of MS2 at pH 9.0 after aggregation at pH = 3.0, supporting the SAXS

Article Snippet: Virus propagation Page 17 of 30 ACS Paragon Plus Environment ACS Nano 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 18 MS2 (DSMZ 1367, DSMZ, Germany) was replicated in Escherichia coli strain W1485 (DSMZ 5695, DSMZ, Germany).

Techniques:

Journal: ACS nano

Article Title: Colloidal Transformations in MS2 Virus Particles: Driven by pH, Influenced by Natural Organic Matter.

doi: 10.1021/acsnano.9b08112

Figure Lengend Snippet: Figure 9: A: Cryo-TEM image of MS2 after disaggregation in presence of NOM at pH 9.0. Single

Article Snippet: Virus propagation Page 17 of 30 ACS Paragon Plus Environment ACS Nano 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 18 MS2 (DSMZ 1367, DSMZ, Germany) was replicated in Escherichia coli strain W1485 (DSMZ 5695, DSMZ, Germany).

Techniques: