Expression profile of MESH1 and ATF4 silencing RCC4 cells

MESH1 encodes for Metazoan SpoT Homolog 1, which is a homologue of bacterial SpoT that mediates bacterial stringent response. In human cells, we found that MESH1-silencing induces an extensive transcriptional response in clear cell carcinoma cell line RCC4 that partially overlap with mammalian cell integrated stress response (ISR), which ATF4 up-regulation is an essential branch of the response. In this study, the goal is to elucidate the role of ATF4 up-regulation in MESH1-silencing transcriptional response. Human clear cell renal carcinoma cell line RCC4 were transfected with siRNA targeting MESH1 (siMESH1), ATF4 (siATF4), or MESH1 with ATF4 together (siMESH1+siATF4), and compared with non-targeting siRNA control (siNT). All samples were prepared as biological triplicates. Total RNA was collected after 3 days of reverse transfection for cDNA synthesis. The influence of silencing MESH1 and ATF4 on gene expression was derived by zero transformation process, in which we compared the gene expression level with siMESH1, siATF4 or siMESH1+siATF4 to the average of gene expression level in control (siNT) samples. biological replicate: siNT-1, siNT-2, siNT-3 biological replicate: siMESH1-1, siMESH1-2, siMESH1-3 biological replicate: siMESH1+siATF-1, siMESH1+siATF-2, siMESH1+siATF-3 biological replicate: siATF4-1, siATF4-2, siATF4-3