Effects of stable overexpression of Tβ10 on cell migration and monolayer wound healing in KKU-M055, KKU-M213 and KKU-M214 sh-T
10-GFP cells. Stable cell lines overexpressing Tβ10 (Lenti-Tβ10) and GFP control plasmid (Lenti-GFP) were generated in KKU-M055 and KKU-M213 cell lines. Selected clones of control and Tβ10 overexpressing KKU-M055 (A) and KKU-M213 (D) cells were confirmed by real time RT-PCR. β-actin was used as an internal control. Arrows indicate selected clone for use in subsequence experiments. Migration potential of KKU-M055 (B) and KKU-M213 (E) cells expressing Lenti-Tβ10 and its controls was determined by the modified Boyden chamber assay at 36 h incubation. Bar graphs represent fold change or percentage of control. n = 3; *P<0.05 versus the control. Wound healing assay was determined in KKU-M055 (C) and KKU-M213 (F). Representative images were taken from the same field at 0, 24, and 48 h in KKU-M055, and 0, 20, and 24 h in KKU-M213 overexpression stable cell line. (G) Rescue experiment in KKU-M214 sh-Tβ10-GFP cells. KKU-M214 sh-Tβ10-GFP cells were transfected with Tβ10 expressing plasmid for 48 h; while the sh-vector-GFP cells received pCMV vector plasmid as a control. The expression levels of Tβ10 in these cells were determined by real time RT-PCR. (H) Cell migration of M214 sh-Tβ10-GFP + Tβ10 plasmid cells and sh-vector-GFP + pCMV cells were determined by the modified Boyden chamber assay at 60 h incubation. Bar graphs represent percentage of control. n = 3; *P<0.05 versus the control. (I) Wound healing assay was determined in M214 sh-Tβ10-GFP + Tβ10 plasmid cells and sh-vector-GFP + pCMV cells. Representative images were taken from the same field at 0, 20, and 24 h.