RNA-Seq experiment of peritoneal macrophgae or IC-21 cell line treated with LPS
A growing number of micro-peptides (miPs) have been identified and annotated in recent years. However, the biological significance of these hidden genomic products remains unclear. In this study, we established a novel strategy to screen human miPs with metabolic relevance by identifying miPs linked to metabolic traits in GWAS datasets. The selected miP, SMIM30, represents a promising drug target due to its capacity to regulate both adipose tissue and systemic insulin sensitivity. We demonstrated that SMIM30 functions as a modulator of inflammatory responses in ex vivo/in vitro macrophage systems. Specifically, isolated mouse peritoneal macrophage and mouse macrophage cell line, IC-21, were treated with LPS with/without SMIM30 expression. Differentially expressed genes can be obtained from RNAseq results, which may guide the characterization of SMIM30-related biological pathways. Overall design: (1) Peritoneal macrophages were isolated from wild-type or SMIM30-Mut mice, with 2´106 cells seeded on the 24-well-plates. Cells were treated with 5ng/ml LPS for 6 hours before collection for RNA isolation. Data shown is triplicates of wild-type or SMIM-30 Mut macrophages. (2) Peritoneal macrophages were isolated from SMIM30-Mut mice, with 2´106 cells seeded on the 24-well-plates. Six hours after seeding, SMIM30-overexpression adenovirus or control adenovirus was added to the wells. Cells were treated with adenovirus for 24h. Cells were then treated with 5ng/ml LPS for 6 hours before collection for RNA isolation. Data shown is triplicates of SMIIM30 overexpression or SMIM-30 Mut macrophages. (3) IC-21 was obtained from ATCC and cultured following manufacturer's instructions. SMIM30 loss-of-function was accomplished by transfection of IC-21 cells with SMIM30 siRNA for 24 hours. Cells were then treated with 5ng/ml LPS for 6 hours before collection for RNA isolation. Data shown is triplicates of wild-type or SMIM-30 knockdown IC-21 cells.