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. 2019 Mar 9;13(5):1002–1017. doi: 10.1002/1878-0261.12439

Figure 5.

Figure 5

The hypomethylating agent 5‐Aza impairs sarcosine‐induced stimulation of Dnmts and cell migration. (A) Immunoblots of Dnmt1, Dnmt3a and Dnmt3b in prostate cells treated with 5‐Aza, sarcosine and their combination (24 h). (B) Densitometry showing quantification of expression of Dnmts normalized to GAPDH. (C) Fold change expression of Dnmts‐encoding mRNA analysed by qRT‐PCR. (D) ICC of DNMT1 in prostate metastatic cells (LNCaP) upon treatment with 5‐Aza, sarcosine and their combination. Scale bar, 30 μm. (E) Cell migration evaluated by monolayer wound‐healing assay. Scale bar, 500 μm. (F) The wounds were quantified at different time points using tscratch software. All values are expressed as the median of five biological replicates (n = 5). (G) Clonogenic assay conducted with the treatment of sarcosine, 5‐Aza and their combination as described in Materials and methods. Left: the full view of wells stained with crystal violet. Right: the quantification of clonal efficiency. The results are represented as the median of five independent experiments (n = 5). The vertical bars indicate the standard deviation. *P < 0.05, **P < 0.01 (paired t‐test).