Nuclear actin assembly is needed for decidualization of human endometrial stromal cells

To know the role of nuclear F-actin in the change of mRNA expression during desidualization, we compared the mRNA expression profiles between cAMP-stimulation and cAMP-stimulation with the overexpression of R62D (actin mutant ActinR62D). Overall design: hESC were overexpressed with mCherry (as a mock control) or R62D, and were treated with or without cAMP for 96 h. RNA extraction was performed by using NucleoSpin® RNA (TaKaRa) and were moved into 1×Reaction buffer from SMART-seq v4 Ultra Low Input RNA Kit (24888N, Takara). SMART-seq library preparation was performed using SMART-seq v4 Ultra Low Input RNA Kit and Nextera DNA Sample Preparation Kit (FC-131-1024, illumine, San Diego, CA) according to the vendor's instruction. Paired-end sequencing (50 bp + 25 bp) was done by using the NextSeq platform (Illumina). Raw reads were first filtered to get rid of low quality reads using Trimmomatic). Reads of less than 20 bases and unpaired reads were also removed. Furthermore, removal of adaptor, polyA, polyT and polyG sequences were performed using Trim Galore! (https://www.bioinformatics.babraham.ac.uk/projects/trim_galore/). For mapping of reads, trimmed reads were first aligned to the human genome hg19 using STAR aligner. Gene counts in triplicate were used to identify differentially expressed genes (DEGs, fulfilling the following criteria: padj<0.05) using DESeq2.