Fig. 3

Differentially expressed miRNAs in IL-4–treated Aps. A The analysis of differentially expressed miRNAs using the small RNA sequencing data of control and IL-4–treated APs (two-day sample). Differentially expressed miRNAs in IL-4–treated APs are marked on the volcano plot as follows, fold change > 1.5 (red) or < -1.5 (blue), p < 0.05. The vertical dashed lines indicate the fold change of 1.5 (black) and 2.0 (blue). B The genomic structure of the H19X-encoded locus. miRNA genes are indicated by pink bars. The genomic region used for transcription factor enrichment analysis is marked with a red line. C The primary transcripts of miR-322, miR-351, and miR-542 were examined by qPCR for the expression levels in APs incubated with IL-4 for 0, 2, and 4 days (n = 3). The let-7b primary transcript was used as a control. D The abundance of mature miRNAs, miR-322-5p, miR-351-5p, and miR-542-3p were examined by qPCR in APs incubated with IL-4 for 0, 2, and 4 days (n = 3). E Transcription factor enrichment analysis for the promoter region of H19X-encoded miRNAs. Top-ranked transcription factors are presented in the table. F Klf4 mRNA levels were measured by qPCR in APs incubated with IL-4 for four days (n = 3). G Klf4 mRNA and primary transcripts of miR-322, miR-503, miR-351, miR-542, and miR-450b were analyzed by qPCR for the expression levels after APs were treated with Klf4 siRNA (n = 3). Data are presented as mean ± SEM. ***p < 0.005, **p < 0.01, *p < 0.05