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. 2021 Jun 30;9:672098. doi: 10.3389/fcell.2021.672098

FIGURE 8.

FIGURE 8

Stable expression of eGFP-vinculin in MCF-7 cells. Following co-transfection of pSB-eGFP-vinculin and pCMV-SB100X cells were cultured in medium containing G418 to allow growth of stably transfected cells only. (A) Graphic illustration of the pSB-eGFP-vinculin plasmid containing the coding sequence of the eGFP-vinculin fusion protein encoded from a CMV promoter. Restrictions used for cloning are indicated. BGH pA, bovine growth hormone polyadenylation site; CMV, cytomegalovirus promoter; IR, internal repeats. (B) Image of eGFP-vinculin in MCF-7 cells subjected to 1g. The white arrows show lamellipodia and vinculin focal adhesion dots. (C) Image of eGFP-vinculin in MCF-7 cells after RPM exposure. The white arrows indicate vinculin dots (D) Image of eGFP-vinculin in MCF-7 MCS following RPM-exposure. Scale bars 10 μm. Expression of mCherry-catenin in MCF-7 cells. MCF-7 cells were transfected with pPBT-mCherry-catenin and images were captured 48 h post transfection. (E) Analysis of the fluorescence intensity of eGFP-vinculin. (F) Graphic illustration of the pSB-eGFP-vinculin plasmid containing the coding sequence of the eGFP-vinculin fusion protein encoded from a CMV promoter. Restrictions used for cloning are indicated. BGH pA, bovine growth hormone polyadenylation site; CMV, cytomegalovirus promoter. (G) Image of MCF-7 cells expressing mCherry-catenin at 1g. (H) Image of RPM-exposed adherent MCF-7 cells expressing mCherry-catenin following incubation at μg. (I) Image of MCF-7 MCS expressing mCherry-catenin following incubation at μg. Scale bars 20 μm. (J) Analysis of the fluorescence intensity of mCherry-catenin. Gene expressions of CTNNB1 (K); CTNNA1 (L); JUN (M); CCND1 (N); NFATC2 (O); BCL9 mRNA (P) and MYC (Q), MCF-7 cells exposed to the RPM. *p < 0.05 1g vs. AD and/or MCS and #p < 0.05 AD vs. MCS.