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. Author manuscript; available in PMC: 2019 Apr 26.
Published in final edited form as: Cancer Res. 2009 Dec 15;69(24):9219–9227. doi: 10.1158/0008-5472.CAN-09-1852

Figure 2. hnRNP A2 knockdown disrupts Golgi morphology and migration of A2780 cells on cell-derived matrix.

Figure 2

(a) A2780 cells transfected with either a control (Con) siRNA, or one targeting hnRNP A2 (A2) were seeded onto cell-derived matrices, and allowed to adhere for approximately 10 hours prior to time lapse microscopy. Frames were captured every 5 minutes over a 500 minute period, and movies generated from these images (Movies S1 and S2). These images are representative stills from the movies. Bar, 50 μm.

(b) Cell tracking software was used to determine cell displacement, migrational persistence, and average speed of migration of A2780 cells on cell-derived matrix following either control (Con) siRNA, or knockdown of hnRNP A2 using two independent oligonucleotides (A2#1 and A2#2). By defining a ‘dither’ as a period of at least 30 minutes in which a cell moves less than 2 μm, the incidence and frequency of cells dithering within a 500 minute period was determined. Also shown is the amount of time during the 500 minute period that the cells spent dithering. Data represent mean ± SEM. * indicates significant difference of p<0.005 and ** indicates p<0.05 (Mann-Whitney test).

(c) A2780 cells were transfected with either a control (Con) or hnRNP A2-targetting siRNA (A2), in combination with a fluorescently-tagged Golgi marker and seeded onto cell-derived matrix. Approximately 10 hours after seeding, timelapse microscopy was used to capture phase-contrast (phase) and fluorescent (Golgi) frames every 5 minutes for a 500 minute time period, and movies were generated from these (Movies S3 and S4). The presented images are representative stills from the movies. Bar, 50 μm.

(d) A2780 cells were transfected with either a control (Con) or hnRNP A2-targetting (A2) siRNA, seeded onto cell-derived matrix, and allowed to adhere for 24 hours before being fixed and stained for γ-tubulin and the Golgi marker, gm130. Bar, 20 μm. The graph shows quantification of proportion of cells with scattered or fragmented Golgi complexes following transfection with either a control (Con) or two hnRNP A2-targetting (A2#1 and A2#2) siRNAs. Data represents mean ± SEM. * indicates statistical significance of p=<0.0001 (Mann-Whitney test).