Journal: Molecular Medicine Reports

Article Title: AQP3 small interfering RNA and PLD2 small interfering RNA inhibit the proliferation and promote the apoptosis of squamous cell carcinoma

doi: 10.3892/mmr.2017.6847

Figure Lengend Snippet: Inhibitory effects of AQP3 and PLD2 siRNA sequences on the target gene. siRNA transfection and reverse transcription-quantitative polymerase chain reaction analyses were performed. The groups were as follows: CK group, normal cultured A431 cells; NC group, A431 cells were transfected with NC siRNA; 1, AQP3-homo-612 siRNA group; 2, AQP3-homo-363 siRNA group; 3, AQP3-homo-360 group; 4, PLD2-homo-602 group; 5, PLD2-homo-1352 group; 6, PLD2-homo-5338 group. siRNA, small interfering RNA; AQP3, aquaporin 3; PLD2, phospholipase D2; CK, control check; NC, negative control.

Article Snippet: The human A431 SCC cell line was purchased from the China Center for Type Culture Collection (Wuhan, China).

Techniques: Transfection, Reverse Transcription, Real-time Polymerase Chain Reaction, Cell Culture, Small Interfering RNA, Control, Negative Control

Journal: Molecular Medicine Reports

Article Title: AQP3 small interfering RNA and PLD2 small interfering RNA inhibit the proliferation and promote the apoptosis of squamous cell carcinoma

doi: 10.3892/mmr.2017.6847

Figure Lengend Snippet: Inhibitory effect of siRNA transfection on the proliferation of A431 cells. Cell proliferation was analyzed and the results are representative of three independent experiments. The statistical significance between two groups was determined using a paired-samples t-test. **P<0.01, compared with the NC group. siRNA, small interfering RNA; AQP3, aquaporin 3; PLD2, phospholipase D2; CK, control check; NC, negative control.

Article Snippet: The human A431 SCC cell line was purchased from the China Center for Type Culture Collection (Wuhan, China).

Techniques: Transfection, Small Interfering RNA, Control, Negative Control

Journal: Molecular Medicine Reports

Article Title: AQP3 small interfering RNA and PLD2 small interfering RNA inhibit the proliferation and promote the apoptosis of squamous cell carcinoma

doi: 10.3892/mmr.2017.6847

Figure Lengend Snippet: AQP3 siRNA and PLD2 siRNA increase the apoptosis of A431 cells. Cell apoptosis was determined; results are representative of three independent experiments. The statistical significance between two groups was determined using a paired-samples t-test. **P<0.01, compared with the NC group. siRNA, small interfering RNA; AQP3, aquaporin 3; PLD2, phospholipase D2; CK, control check; NC, negative control.

Article Snippet: The human A431 SCC cell line was purchased from the China Center for Type Culture Collection (Wuhan, China).

Techniques: Small Interfering RNA, Control, Negative Control

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