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. 2021 Sep 15;114(2):290–301. doi: 10.1093/jnci/djab183

Figure 5.

Figure 5.

Regulation of Serglycin (SRGN) gene expression by DNA methylation. Immunoblot analyses of SRGN expression in the conditioned media from H2030 and DFCI024 cell lines A) after TTF-1 overexpression and B) treated with negative control siRNA or NRF2 siRNA. Experiments have been performed with at least 3 independent biological repeats. C) Hierarchical clustering analysis of the top 15% methylated CpG sites (n = 4137) of an Illumina Infinium HumanMethylation27 BeadChip microarray in 35 LUAD cell lines. P values were calculated using Fisher’s exact test. C1 and C2, clusters 1 and 2. D) Correlation of SRGN mRNA expression and the methylation levels of the 5 CpG sites in the SRGN promoter region in 41 LUAD cell lines. Correlation coefficients and P values were calculated using Spearman correlation test. To adjust P values for multiple comparisons, the Bonferroni correction was conducted. E) Quantitative reverse transcription polymerase chain reaction (RT-PCR) analysis of SRGN mRNA expression levels (left) and the methylation levels of the 5 CpG sites in the SRGN promoter region (right) after treatment with 5-Aza-dC at 2 µM in H3255 and H920 cell lines. F) Quantitative RT-PCR analysis of DNA methyltransferase 1 (DNMT1), DNMT3A, DNMT3B, and SRGN mRNA expression levels in H920 cells treated with negative control siRNA or siRNA against DNMT1, DNMT3A, or DNMT3B. In E and F, experiments have been performed with at least 3 independent biological repeats. mRNA expression was presented as the average and SD of triplicate samples from a representative experiment. All statistical tests were 2-sided.