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Fig. 1 | Breast Cancer Research

Fig. 1

From: Calmodulin-like protein 3 is an estrogen receptor alpha coregulator for gene expression and drug response in a SNP, estrogen, and SERM-dependent fashion

Fig. 1

CALML3 as a potential ERα coregulator binds to the ZNF423 rs9940645 SNP. a Schematic representation of the genomic region surrounding the rs9940645 SNP (in intron 1) in the ZNF423 gene. The SNP is in intron 2 of ZNF423 and 240 bp away from the closest ERE. b EMSA to identify the candidate proteins that bind differentially to WT and variant SNPs sequences in LCLs treated with E2 ± 4-OH-TAM. 13 bands indicating specific DNA–protein binding. c Further validation of CALML3 as a candidate protein by ChIP-qPCR using ERα antibody after the knockdown of CALML3 in WT and variant LCLs treated with E2 ± 4-OH-TAM. Primers used for amplification are indicated in (a) with arrows, so the amplicons included both the SNP and EREs. YWHAZ as a nonspecific SNP-binding protein identified by MS was also included as a negative control. Enrichment of DNA fragments shown as the mean (± SEM) of three independent experiments. *p < 0.05, **p < 0.01 by two-tailed Student’s t test. 4-OH-TAM 4-hydroxytamoxifene, E2 estradiol, ERα estrogen receptor alpha, ERE estrogen response element, ns not significant, WT wild type

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