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Fig. 3 | BMC Evolutionary Biology

Fig. 3

From: Tinkering signaling pathways by gain and loss of protein isoforms: the case of the EDA pathway regulator EDARADD

Fig. 3

The two isoforms of EDARADD are expressed by two differentially regulated alternative promoters. a Scheme of the promoter constructs. The human promoter A region has no homologous region in mouse. Homo, Homo sapiens; Mus, Mus musculus. b In cellulo test of promoters activity. Constructs shown in A were transfected in HEK cells. Error bars show variation between triplicates of one out of 3 experiments. Y axis: normalized luciferase activity against β-galactosidase activity. c In cellulo test of promoters activity. B constructs shown in B were transfected in HEK cells. Three independent experiments were performed and yielded similar results. One out of these three experiments is shown with error bars showing variation between triplicates within this given experiment. Y axis: normalized luciferase activity against β-galactosidase activity. d Promoters response to EDA pathway activation by EDAR co-transfection. Constructs shown in A were co-transfected with increasing amount of an EDAR expressing plasmid: 0, 50, 100 or 200 ng. Error bars show variation between triplicates in one out of 3 experiments that yielded similar results. Y axis: normalized luciferase activity against β-galactosidase activity. e Promoters response to EDA pathway activation by EDAR co-transfection. B constructs were co-transfected with increasing amount of an EDAR expressing plasmid: 0, 50, 100 or 200 ng. Error bars show variation between triplicates in one out of 3 experiments that yielded similar results. Y axis: normalized luciferase activity against β-galactosidase activity. f Promoter B response to different NF-κB modulators. The human B promoter was co-transfected with 200 ng of TRAF 6, 200 ng of EDAR only or 200 ng of EDAR and 200 ng of CYLD supplemented or not by empty vector. Y axe: normalized luciferase activity as in B. g Expression of the two EDARADD isoforms. Expression of the two isoforms were assayed by RT-PCR in different tissues in human (Homo sapiens), a rodent guinea pig (Cavia porcellus) and a marsupial tammar wallaby (Macropus eugenii) (AB: A and B detected; B: only B detected; na: not assessed) and in different human cell lines using ENCODE RNAseq data (A = B : A and B co-expressed at similar levels; B > A: B expressed at higher levels; B: only B is detected). Cell lines: GM92, NHEK, GM78, H1ES, K562, MCF7, GM91

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