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Generation of PCDHGC3 knockout in breast cancer and melanoma cell lines. Western blot analysis of wild type (WT), control (C; transfected with the PCDH2 HDR Plasmid (h2) containing a puromycin resistance gene) and knockout (KO) HCC1806 breast cancer cell line ( a ) and A2058 melanoma cell line ( b ). β-Actin served as an endogenous control.

Journal: NeuroSci

Article Title: The Knockout of Protocadherin Gamma C3 (PCDHGC3) in Breast Cancer and Melanoma Cell Lines Leads to Increased Adhesion of Knockout Cells to Brain Microvascular Endothelial Cells

doi: 10.3390/neurosci7020047

Figure Lengend Snippet: Generation of PCDHGC3 knockout in breast cancer and melanoma cell lines. Western blot analysis of wild type (WT), control (C; transfected with the PCDH2 HDR Plasmid (h2) containing a puromycin resistance gene) and knockout (KO) HCC1806 breast cancer cell line ( a ) and A2058 melanoma cell line ( b ). β-Actin served as an endogenous control.

Article Snippet: Melanoma cell line A2058 (CRL-3601, ATCC, Manassas, VA, USA) was cultured in Minimum Essential Medium Eagle (MEM) (M5650) containing 2% MEM Non-essential-Amino Acid Solution (M7145, Sigma-Aldrich, St. Louis, MO, USA), 10% FCS, L-glutamine and penicillin/streptomycin.

Techniques: Knock-Out, Western Blot, Control, Transfection, Plasmid Preparation

Cell proliferation assay in control and PCDHGC3 knockout cells. Proliferation rate of control and PCDHGC3 knockout (KO) HCC1806 breast cancer cells ( a ) and of control and PCDHGC3 KO A2058 melanoma cells ( b ). **** = p < 0.0001, unpaired t test.

Journal: NeuroSci

Article Title: The Knockout of Protocadherin Gamma C3 (PCDHGC3) in Breast Cancer and Melanoma Cell Lines Leads to Increased Adhesion of Knockout Cells to Brain Microvascular Endothelial Cells

doi: 10.3390/neurosci7020047

Figure Lengend Snippet: Cell proliferation assay in control and PCDHGC3 knockout cells. Proliferation rate of control and PCDHGC3 knockout (KO) HCC1806 breast cancer cells ( a ) and of control and PCDHGC3 KO A2058 melanoma cells ( b ). **** = p < 0.0001, unpaired t test.

Article Snippet: Melanoma cell line A2058 (CRL-3601, ATCC, Manassas, VA, USA) was cultured in Minimum Essential Medium Eagle (MEM) (M5650) containing 2% MEM Non-essential-Amino Acid Solution (M7145, Sigma-Aldrich, St. Louis, MO, USA), 10% FCS, L-glutamine and penicillin/streptomycin.

Techniques: Proliferation Assay, Control, Knock-Out

Relative adhesion of PCDHGC3 knockout breast cancer and melanoma cells to human in vitro BBB models. Adhesion measurements of HCC1806 PCDHGC3 knockout (KO) and control cells to hCMEC/D3 ( a ) and BLECs ( b ) after 30, 60, and 120 min. Adhesion measurements of A2058 PCDHGC3 knockout (KO) and control cells to hCMEC/D3 ( c ) and BLECs ( d ) after 30, 60, and 120 min. Control cell adhesion was measured at each time point; however, for clarity, only the measurement after 30 min is shown. Data are presented as mean relative adhesion versus control with standard deviation, **** = p ≤ 0.0001, one-way ANOVA test.

Journal: NeuroSci

Article Title: The Knockout of Protocadherin Gamma C3 (PCDHGC3) in Breast Cancer and Melanoma Cell Lines Leads to Increased Adhesion of Knockout Cells to Brain Microvascular Endothelial Cells

doi: 10.3390/neurosci7020047

Figure Lengend Snippet: Relative adhesion of PCDHGC3 knockout breast cancer and melanoma cells to human in vitro BBB models. Adhesion measurements of HCC1806 PCDHGC3 knockout (KO) and control cells to hCMEC/D3 ( a ) and BLECs ( b ) after 30, 60, and 120 min. Adhesion measurements of A2058 PCDHGC3 knockout (KO) and control cells to hCMEC/D3 ( c ) and BLECs ( d ) after 30, 60, and 120 min. Control cell adhesion was measured at each time point; however, for clarity, only the measurement after 30 min is shown. Data are presented as mean relative adhesion versus control with standard deviation, **** = p ≤ 0.0001, one-way ANOVA test.

Article Snippet: Melanoma cell line A2058 (CRL-3601, ATCC, Manassas, VA, USA) was cultured in Minimum Essential Medium Eagle (MEM) (M5650) containing 2% MEM Non-essential-Amino Acid Solution (M7145, Sigma-Aldrich, St. Louis, MO, USA), 10% FCS, L-glutamine and penicillin/streptomycin.

Techniques: Knock-Out, In Vitro, Control, Standard Deviation

PCDHGC3 KO leads to stronger invasion of PCDHGC3 knockout breast cancer and melanoma cells. HCC1806 PCDHGC3 knockout (KO) and control cells ( a ) and A2058 PCDHGC3 knockout (KO) and control ( b ) invaded for 48 h through Transwells coated with Matrigel. The number of invaded cells is shown. Data are presented as mean cell number with standard deviation, * = p ≤ 0.05, unpaired t -test.

Journal: NeuroSci

Article Title: The Knockout of Protocadherin Gamma C3 (PCDHGC3) in Breast Cancer and Melanoma Cell Lines Leads to Increased Adhesion of Knockout Cells to Brain Microvascular Endothelial Cells

doi: 10.3390/neurosci7020047

Figure Lengend Snippet: PCDHGC3 KO leads to stronger invasion of PCDHGC3 knockout breast cancer and melanoma cells. HCC1806 PCDHGC3 knockout (KO) and control cells ( a ) and A2058 PCDHGC3 knockout (KO) and control ( b ) invaded for 48 h through Transwells coated with Matrigel. The number of invaded cells is shown. Data are presented as mean cell number with standard deviation, * = p ≤ 0.05, unpaired t -test.

Article Snippet: Melanoma cell line A2058 (CRL-3601, ATCC, Manassas, VA, USA) was cultured in Minimum Essential Medium Eagle (MEM) (M5650) containing 2% MEM Non-essential-Amino Acid Solution (M7145, Sigma-Aldrich, St. Louis, MO, USA), 10% FCS, L-glutamine and penicillin/streptomycin.

Techniques: Knock-Out, Control, Standard Deviation

Relative expression of target genes in PCDHGC3 KO breast cancer and melanoma cells. The relative expression (RQ value) of each target in PCDHGC3 KO HCC1806 ( a ) and PCDHGC3 KO A2058 ( b ) cells relative to control cells is shown. A RQ value < 1.0 indicates decreased expression, a RQ value > 1.0 indicates increased expression compared to the control cells. The means with standard deviation are shown as the fold of the control. * = p ≤ 0.05, ** = p ≤ 0.01, *** = p ≤ 0.001, **** = p ≤ 0.0001, unpaired t -test.

Journal: NeuroSci

Article Title: The Knockout of Protocadherin Gamma C3 (PCDHGC3) in Breast Cancer and Melanoma Cell Lines Leads to Increased Adhesion of Knockout Cells to Brain Microvascular Endothelial Cells

doi: 10.3390/neurosci7020047

Figure Lengend Snippet: Relative expression of target genes in PCDHGC3 KO breast cancer and melanoma cells. The relative expression (RQ value) of each target in PCDHGC3 KO HCC1806 ( a ) and PCDHGC3 KO A2058 ( b ) cells relative to control cells is shown. A RQ value < 1.0 indicates decreased expression, a RQ value > 1.0 indicates increased expression compared to the control cells. The means with standard deviation are shown as the fold of the control. * = p ≤ 0.05, ** = p ≤ 0.01, *** = p ≤ 0.001, **** = p ≤ 0.0001, unpaired t -test.

Article Snippet: Melanoma cell line A2058 (CRL-3601, ATCC, Manassas, VA, USA) was cultured in Minimum Essential Medium Eagle (MEM) (M5650) containing 2% MEM Non-essential-Amino Acid Solution (M7145, Sigma-Aldrich, St. Louis, MO, USA), 10% FCS, L-glutamine and penicillin/streptomycin.

Techniques: Expressing, Control, Standard Deviation

Matrix metalloproteinase (MMP) activity in cell culture medium of PCDHGC3 knockout (KO) breast cancer and melanoma cells. Fluorescence signal of MMP-substrate (SB) cleavage in the cell culture medium of PCDHGC3 KO HCC1806 ( a ) and PCDHGC3 KO A2058 ( b ) cells and control cells is expressed as relative fluorescence units (RFU) ± standard deviation. ** = p < 0.01, *** = p < 0.001, unpaired t test.

Journal: NeuroSci

Article Title: The Knockout of Protocadherin Gamma C3 (PCDHGC3) in Breast Cancer and Melanoma Cell Lines Leads to Increased Adhesion of Knockout Cells to Brain Microvascular Endothelial Cells

doi: 10.3390/neurosci7020047

Figure Lengend Snippet: Matrix metalloproteinase (MMP) activity in cell culture medium of PCDHGC3 knockout (KO) breast cancer and melanoma cells. Fluorescence signal of MMP-substrate (SB) cleavage in the cell culture medium of PCDHGC3 KO HCC1806 ( a ) and PCDHGC3 KO A2058 ( b ) cells and control cells is expressed as relative fluorescence units (RFU) ± standard deviation. ** = p < 0.01, *** = p < 0.001, unpaired t test.

Article Snippet: Melanoma cell line A2058 (CRL-3601, ATCC, Manassas, VA, USA) was cultured in Minimum Essential Medium Eagle (MEM) (M5650) containing 2% MEM Non-essential-Amino Acid Solution (M7145, Sigma-Aldrich, St. Louis, MO, USA), 10% FCS, L-glutamine and penicillin/streptomycin.

Techniques: Activity Assay, Cell Culture, Knock-Out, Fluorescence, Control, Standard Deviation