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. 2021 Apr 27;12(5):650. doi: 10.3390/genes12050650

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Analysis of FOXC2 wild-type and mutant proteins after FOXC2-AS1 overexpression or silencing. (A) Western blot analysis of FOXC2 protein expression in HeLa cells co-transfected with different expression vectors and siRNAs (GFP-FOXC2wt; GFP-FOXC2wt + siNC; GFP-FOXC2wt + FOXC2-AS1; GFP-FOXC2wt + siAS1; GFP-FOXC2(H199Pfs264*); GFP-FOXC2(H199Pfs264*) + siNC; GFP-FOXC2(H199Pfs264*) + FOXC2-AS1; GFP-FOXC2(H199Pfs264*) + siAS1). Lane 1: non-transfected HeLa cells. (B) Immunofluorescence analysis of HeLa cells after transfection with GFP-FOXC2wt; GFP-FOXC2wt + FOXC2-AS1; GFP-FOXC2wt + siAS1; GFP-FOXC2(H199Pfs264*); GFP-FOXC2(H199Pfs264*) + FOXC2-AS1; GFP-FOXC2(H199Pfs264*) + siAS1. After 48 h, the cells were fixed with 3% paraformaldehyde and FOXC2 recombinant proteins were observed by direct immunofluorescence analysis of GFP tag (in green); 20× magnification. Scale bar: 10 μm.