Fig 1: Knockdown of ACTN1 inhibits growth and metastasis of HNSCC in vivo. (A and B) The appearances of mice and tumor in the two groups. (C) Mice weight changed from day 0 to day 21. (D) Tumor weight in mice transfected with shRNA-ACTN1 was determined. (E) Tumor volume changed from day 0 to day 21. (F) The expression of Ki-67 in tumor tissues was analyzed by immunohistochemistry staining. (G and H) ITGA5 mRNA and protein expression in mice transfected with shRNA-ACTN1 were detected by RTqPCR and Western blot analysis. *P<0.05, **P<0.01 and ***P<0.001 vs. shRNA-NC group
Fig 2: Knockdown of ACTN1 inhibits invasion, migration and EMT of HNSCC cells
Fig 3: Knockdown of ACTN1 inhibits proliferation of HNSCC cells. (A and B) ACTN1 mRNA and protein expression in Tu686 cells transfected with shRNA-ACTN1 were detected by RTqPCR and Western blot analysis. ***P<0.001 vs. Control group. ###P<0.001 vs. shRNA-NC group. ∆P<0.05 vs. shRNA-ACTN1#1 group. The viability (C), proliferation (D) and colony formation ability (E) of Tu686 cells transfected with shRNA-ACTN1 were detected by CCK-8 assay, EdU staining and colony formation assay respectively. (F) The expression of proliferation related proteins in Tu686 cells transfected with shRNA-ACTN1 was detected by Western blot analysis. **P<0.01 and ***P<0.001 vs. Control group. ##P<0.01 and ###P<0.001 vs. shRNA-NC group
Fig 4: ACTN1 is highly expressed in HNSCC and is associated with poor prognosis. (A) ACTN1 expression in HNSCC tissues from GEPIA database. (B) The relation between ACTN1 expression and overall survival. (C and D) ACTN1 mRNA and protein expression in HNSCC cells transfected with shRNA-ACTN1 were detected by RTqPCR and Western blot analysis. *P<0.05 and ***P<0.001 vs. HIOEC group
Fig 5: Overexpression of ITGA5 reverses the inhibitory effect of ACTN1 knockdown on the proliferation of HNSCC cells. (A and B) ITGA5 mRNA and protein expression in Tu686 cells transfected with ov-ITGA5 were detected by RTqPCR and Western blot analysis. *P<0.05 and ***P<0.001 vs. Control group. #P<0.05 and ###P<0.001 vs. shRNA-NC group. ∆P<0.05 vs. shRNA-ACTN1#1 group. The viability (C), proliferation (D) and colony formation ability (E) of Tu686 cells transfected with shRNA-ACTN1 and ov-ITGA5 were detected by CCK-8 assay, EdU staining and colony formation assay, respectively. (F) The expression of proliferation-related proteins in Tu686 cells transfected with shRNA-ACTN1 and ov-ITGA5 was detected by Western blot analysis. **P<0.01 and ***P<0.001 vs. shRNA-NC group. ##P<0.01 and ###P<0.001 vs. shRNA-ACTN1+ov-NC group
Supplier Page from Abcam for Anti-alpha Actinin/ACTN1 antibody