Skip to main content
Fig. 5 | Cell Communication and Signaling

Fig. 5

From: The role of the redox/miR-6855-3p/PRDX5A axis in reversing SLUG-mediated BRCA2 silencing in breast cancer cells

Fig. 5

Nuclear PRDX5 originates from the second in-frame ATG codon in the PDRX5 ORF. a Schematics of the PRDX5A constructs in the p3XFLAG-CMV 14 vector. The wildtype ORF with ATG1 and ATG2 was cloned to express a 24-kDa protein FLAG-tagged at the C-terminus. Constructs with mutations at the start codons (ATG1-mutant and ATG2-mutant) were also generated. CP and CR represent codons for active site Cys. MLS, mitochondrial localization signal; NLS, nuclear localization signal; 3X-FLAG, FLAG tag from the vector. b Immunoblot showing expression of the recombinant proteins. The wildtype and ATG2-mutant (both ~ 24 kDa, a) existed as precursor proteins. The size of the mature form (~ 18 kDa, b) is the same as that of the protein expressed by the ATG2-mutant construct. Fibrillarin and GSK3β served as a nuclear marker and a loading control, respectively. c Western blot quantification for the levels nuclear PRDX5A recombinant proteins. Results are mean ± SE (n = 3). The difference is statistically significant (p < 0.0001). d Immunofluorescence analysis showing nuclear localization of FLAG-tagged PRDX5A (wildtype), ATG1-mutant, and ATG2-mutant in unsynchronized cells. Anti-FLAG M2 antibody was used to detect FLAG-tagged PRDX5A (red) and DAPI was used as a nuclear stain (blue)

Back to article page