Journal: Protein Engineering, Design and Selection

Article Title: Discovery of internalizing antibodies to basal breast cancer cells

doi: 10.1093/protein/gzx063

Figure Lengend Snippet: Characterization of human mAbs targeting EphA2 and CD73. (A) 2D6 (EphA2) and 2B4 (CD73) IgG antibodies reduced the colony formation of MDA-MB-231 cells in a soft agar colony forming assay. (B) 2D6 reduced the surface level of EphA2 on MDA-MB-231 cells, while the commercial anti-EphA2 MAb D7 and the anti-EGFR 2224-IgG had no effect. (C) 2D6 inhibited MDA-MB-231 cells invasion in a modified Boyden Chamber Assay with the ligand Ephrin A1 as a control

Article Snippet: Modified Boyden chambers (8-μm pore size, Transwell, BD-Falcon) were used in cell-invasion assays as described previously ( Lochter et al. , 1997 ).

Techniques: Modification, Boyden Chamber Assay, Control

Journal: Frontiers in Oncology

Article Title: Effect of CRISPR Knockout of AXIN1 or ARID1A on Proliferation and Migration of Porcine Hepatocellular Carcinoma

doi: 10.3389/fonc.2022.904031

Figure Lengend Snippet: AXIN1 knockout increases the migration, but does not alter the proliferation, of porcine HCC cells. (A, B) Cell proliferation was determined for A272 (A) and A274 (B) parental and AXIN1 KO cells by MTS assay. Cell viability at different time points (days) was measured and expressed as A 490 normalized to values of day 1. Statistical analysis was conducted to compare viability of the different cell lines at each time point by two-way ANOVA. No significant difference was detected between the cell lines at p < 0.05. (C, D) Migration of A272 AXIN1 KO (C) and A274 AXIN1 KO (D) cells in comparison to parental cells was assessed by transwell cell migration assay. Quantitated migration ability is presented as the number of migrated cells per field. Values in the bar graph represent mean ± S.E. (n = 6 fields). ***, p < 0.0001. Representative images of migrated cells stained with crystal violet are shown below each bar graph. Scale bar, 250 μm.

Article Snippet: Cell migration was analyzed using a modified two chamber transwell system (#353097; Corning, Corning, NY, USA).

Techniques: Knock-Out, Migration, MTS Assay, Comparison, Cell Migration Assay, Staining

Journal: Frontiers in Oncology

Article Title: Effect of CRISPR Knockout of AXIN1 or ARID1A on Proliferation and Migration of Porcine Hepatocellular Carcinoma

doi: 10.3389/fonc.2022.904031

Figure Lengend Snippet: ARID1A knockout increases the proliferation and migration of porcine HCC cells. (A, B) Cell proliferation was determined for A272 (A) and A274 (B) parental and ARID1A KO cells by MTS assay. Cell viability at different time points (days) was measured and expressed as A 490 normalized to values of day 1. Statistical analysis was conducted to compare viability of the different cell lines at each time point by two-way ANOVA. *, p < 0.05; **, p < 0.005. (C, D) Migration of A272 ARID1A KO (C) and A274 ARID1A KO (D) cells in comparison to parental cells was assessed by transwell cell migration assay. Quantitated migration ability is presented as the number of migrated cells per field. Values in the bar graph represent mean ± S.E. (n = 6 fields). ***, p < 0.0001. Representative images of migrated cells stained with crystal violet are shown below each bar graph. Scale bar, 250 μm.

Article Snippet: Cell migration was analyzed using a modified two chamber transwell system (#353097; Corning, Corning, NY, USA).

Techniques: Knock-Out, Migration, MTS Assay, Comparison, Cell Migration Assay, Staining

Journal: Frontiers in Oncology

Article Title: Effect of CRISPR Knockout of AXIN1 or ARID1A on Proliferation and Migration of Porcine Hepatocellular Carcinoma

doi: 10.3389/fonc.2022.904031

Figure Lengend Snippet: Simultaneous knockout of ARID1A and AXIN1 in Oncopig HCC cells increases the migration, but does not alter the proliferation of porcine HCC cells. (A) CRISPR/Cas9-induced editing of AXIN1 and ARID1A in A272 and A274 porcine HCC cells using AXIN1 gRNA#2 and ARID1A gRNA. Genomic DNA was collected two days post-transfection and analyzed by targeted NGS. The bar graph depicts the percentages (%) of total reads that displayed indels at the gRNA target site. (B, C) Top 10 reads detected by targeted NGS analysis in the two porcine HCC lines mapped to the reference sequence. The percentage of reads of each sequence are shown on the right. The asterisk (*) indicates/marks non-edited reads.. Dashed line, predicted Cas9 cleavage position; red box, insertion; dash, deleted base. (D) NGS Analysis of ARID1A-AXIN1 KO single cell clone isolated from A272 porcine HCC cells transfected with Cas9, AXIN1 gRNA#2, and ARID1A gRNA. Dashed line, predicted Cas9 cleavage position; red box, insertion; dash, deleted base. (E) Confirmation of complete loss of AXIN1 protein in A272 ARID1A-AXIN1 KO porcine HCC cells by Western blotting. The cells were lysed two days post-transfection and analyzed using an anti-AXIN1 antibody. β-actin was used as a loading control. (F) Cell proliferation was determined for A272 parental and ARID1A-AXIN1 KO cells by MTS assay. Cell viability at different time points (days) was measured and expressed as A 490 normalized to values of day 1. Statistical analysis was conducted to compare viability of the different cell lines at each time point by two-way ANOVA. No significant difference was detected between the cell lines at p < 0.05. (G) Migration of A272 ARID1A-AXIN1 KO cells in comparison to parental cells was assessed by transwell cell migration assay. Quantitated migration ability is presented as the number of migrated cells per field. Values in the bar graph represent mean ± S.E. (n = 6 fields). ***, p < 0.0001. Representative images of migrated cells stained with crystal violet are shown below each bar graph. Scale bar, 250 μm.

Article Snippet: Cell migration was analyzed using a modified two chamber transwell system (#353097; Corning, Corning, NY, USA).

Techniques: Knock-Out, Migration, CRISPR, Transfection, Sequencing, Isolation, Western Blot, Control, MTS Assay, Comparison, Cell Migration Assay, Staining

Journal: Journal of Cancer

Article Title: A Luminacin D Analog HL142 Inhibits Ovarian Tumor Growth and Metastasis by Reversing EMT and Attenuating the TGFβ and FAK Pathways

doi: 10.7150/jca.61066

Figure Lengend Snippet: HL142 inhibits proliferation, colony formation, migration and invasion in OC cells. A. Cell colonies in OVCAR3 and OVCAR8 cells following 10 µM HL142-treatment for two weeks and stained with Crystal Violet. B. MTT analysis of cell proliferation in OVCAR3 and OVCAR8 cells treated with different doses of HL142 at different time points. C. Transwell cell migration assay of OVCAR3 and OVCAR8 cells treated with 10μM HL142 for 24h and migrated cells were stained with Crystal Violet. D. Matrigel-coated transwell invasion assay of OVCAR3 and OVCAR8 cells treated with 10μM HL142 for 48h and stained with H&E (*** p <0.001, ** p <0.01, * p <0.05).

Article Snippet: Cell migration was analyzed using modified Transwell chambers (BD FalconTM, San Jose, CA) inserted into 24-well culture plates.

Techniques: Migration, Staining, MTT Cell Proliferation, Cell Migration Assay, Transwell Invasion Assay