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. 2014 May 23;9(5):e98208. doi: 10.1371/journal.pone.0098208

Figure 6. Talin-1 knockdown decreased cell adhesion, migration and invasion of SCC-9, A431 and SCC-9 LN1 cells.

Figure 6

(a) Talin-1 showed lower protein expression in SCC-9/siRNA TLN-1 cells compared to SCC-9/control (scrambled) by immunoblotting. The total proteins (30 µg) were submitted to 1-D electrophoresis on 12% SDS-polyacrylamide gels, they were transferred onto nitrocellulose membrane and incubated with anti-talin-1 antibody. Anti-GAPDH antibody was used as loading control. The graph represents the normalized optical density. (b) Talin-1 mRNA expression levels in SCC-9/siRNA TLN-1 cells compared to SCC-9/control (scrambled) by qRT-PCR (n = 3, Student's t-test, p<0.05). The data were normalized with GAPDH gene. (c) SCC-9/control (scrambled) and SCC-9/siRNA TLN-1 cells, A431/control (scrambled) and A431/siRNA TLN-1 cells, SCC-9 LN1/control (scrambled) and SCC-9 LN1/siRNA TLN-1 cells were seeded in Matrigel coated 96-well plates. After 1 h, cells were stained and the cell adhesion was measured (n = 3, * indicates p<0.05, Student's t-test for each comparison). (d) SCC-9/control (scrambled) and SCC-9/siRNA TLN-1 cells, A431/control (scrambled) and A431/siRNA TLN-1 cells, SCC-9 LN1/control (scrambled) and SCC-9 LN1/siRNA TLN-1 cells were seeded in serum-free media in the upper chamber of transwell plates and were allowed to migrate towards the lower chamber containing 1% FBS supplemented media (n = 3, * indicates p<0.05, Student's t-test). (e) SCC-9/control (scrambled) and SCC-9/siRNA TLN-1 cells, A431/control (scrambled) and A431/siRNA TLN-1 cells, SCC-9 LN1/control (scrambled) and SCC-9 LN1/siRNA TLN-1 cells were seeded in serum-free media in the upper chamber of matrigel-coated transwell plates and were allowed to invade towards the lower chamber containing 10% FBS supplemented media (n = 2, * indicates p<0.05, Student's t-test).