Figure 8
From: G-protein-coupled receptor participates in 20-hydroxyecdysone signaling on the plasma membrane
ChIP analysis showing ErGPCR regulates 20E-induced EcRB1 binding to EcRE through cellular Ca2+signaling. (A) ErGPCR knockdown suppresses the EcRB1 binding to EcRE under 20E treatment. Cells were transfected with pIEx-4-RFP or pIEx-4-EcRB1-RFP for 24 h. The cells were treated by dsErGPCR or dsGFP for 12 h, then exposed in 1 μM 20E or DMSO for 6 h. (B) FL and Pyr3 repress EcRB1 binding to EcRE in 20E treatment. Cells were transfected with pIEx-4-RFP or pIEx-4-EcRB1-RFP for 48 h. The cells were subjected in 1 μM 20E or DMSO for 6 h after different calcium channel inhibitors pretreatment for 1 h. ChIP was carried out without antibody (negative control) or with anti-RFP antibody. The DNA fragment (EcRE) was amplified by qRT-PCR using HHR3F/R primers. Input is the positive control of nonimmunoprecipitated chromatin.