DOI: 10.5061/dryad.2330r1b
title: Data from: Vasohibin1, a new IRES trans-acting factor for induction of (lymph)angiogenic factors in early hypoxia

description: Hypoxia, a major inducer of angiogenesis, triggers major changes of gene
 expression at the transcriptional level. Furthermore, global protein
 synthesis is blocked while internal ribosome entry sites (IRES) allow
 specific mRNAs to be translated. Here we report the transcriptome and
 translatome signatures of (lymph)angiogenic genes in hypoxic HL-1
 cardiomyocytes: most genes are not induced at the transcriptome-, but at
 the translatome level, including all IRES-containing mRNAs. Our data
 reveal activation of (lymph)angiogenic factor mRNA IRESs in early hypoxia.
 We identify vasohibin1 (VASH1) as an IRES trans-acting factor (ITAF) able
 to bind RNA and to activate FGF1 and VEGFD IRESs in hypoxia while it
 exhibits an inhibitory effect in normoxia. This suggests a generalized
 process of IRES-dependent induction of (lymph)angiogenic growth factors in
 early hypoxia, with a variable IRESome composition. IRES-dependent
 translation thus appears as a pivotal process to trigger new vessel
 formation in ischemic heart.
description: pTRIP-CRF1AL+
 bicistronic lentivector with FGF1 IRESThe bicitronic
 lentivector plasmid codes for renilla luciferase LucR and firefly
 luciferase Luc+ under the control of the cytomegalovirus (CMV) promoter,
 with the human FGF1 mRNA IRES A (430 nt-long leader A, Martineau et al,
 Mol Cell Biol. 2004) between the two reporter genes. The LucR gene used
 here is a modified version of LucR where all the predicted splice donor
 sites have been mutated. pTRIP is a Delta-U3 plasmid allowing to produce
 self-inactivating (SIN) lentivectors.pTRIP-CRF1AL+.apepTRIP-CRF1AL+.apepTRIP-CRFL+ bicistronic lentivector with FGF2 IRESThe bicitronic lentivector plasmid codes for renilla luciferase LucR and firefly luciferase Luc+ under the control of the cytomegalovirus (CMV) promoter, with the human FGF2 mRNA IRES (480 nt-long leader region, Vagner et al, Mol Cell Biol. 1995) between the two reporter genes. The LucR gene used here is a modified version of LucR where all the predicted splice donor sites have been mutated. pTRIP is a Delta-U3 plasmid allowing to produce self-inactivating (SIN) lentivectors.pTRIP-CRFL+.apepTRIP-CRFL+.apepTRIP-CRVAaL+ bicistronic lentivector with VEGFA IRES aThe bicitronic lentivector plasmid codes for renilla luciferase LucR and firefly luciferase Luc+ under the control of the cytomegalovirus (CMV) promoter, with the human VEGFA mRNA 302 nt-long IRES a between the two reporter genes (Huez et al, Mol Cell Biol. 1998). The LucR gene used here is a modified version of LucR where all the predicted splice donor sites have been mutated. pTRIP is a Delta-U3 plasmid allowing to produce self-inactivating (SIN) lentivectors.pTRIP-CRVAaL+.apepTRIP-CRVAaL+.apepTRIP-CRVAaL+.apepTRIP-CRVAbL+ bicistronic lentivector with VEGFA IRES bThe bicitronic lentivector plasmid codes for renilla luciferase LucR and firefly luciferase Luc+ under the control of the cytomegalovirus (CMV) promoter, with the human VEGFA 485 nt-long IRES b (Huez et al, Mol Cell Biol. 1998) between the two reporter genes. The LucR gene used here is a modified version of LucR where all the predicted splice donor sites have been mutated. pTRIP is a Delta-U3 plasmid allowing to produce self-inactivating (SIN) lentivectors.pTRIP-CRVAbL+.apepTRIP-CRVAbL+.apepTRIP-CRhVCL+ bicistronic lentivector with VEGFC IRESThe bicitronic lentivector plasmid codes for renilla luciferase LucR and firefly luciferase Luc+ under the control of the cytomegalovirus (CMV) promoter, with the human VEGFC 419 nt-long IRES (Morfoisse et al, Cell Reports 2014) between the two reporter genes. The LucR gene used here is a modified version of LucR where all the predicted splice donor sites have been mutated. pTRIP is a Delta-U3 plasmid allowing to produce self-inactivating (SIN) lentivectorspTRIP-CRhVCL+.apepTRIP-CRhVCL+.apepTRIP-CRhVDL+ bicistronic lentivector with VEGFD IRESThe bicitronic lentivector plasmid codes for renilla luciferase LucR and firefly luciferase Luc+ under the control of the cytomegalovirus (CMV) promoter, with the human VEGFD 507 nt-long IRES (Morfoisse et al, Cancer Res 2016) between the two reporter genes. The LucR gene used here is a modified version of LucR where all the predicted splice donor sites have been mutated. pTRIP is a Delta-U3 plasmid allowing to produce self-inactivating (SIN) lentivectorspTRIP-CRhVDL+.apepTRIP-CRhVDL+.apepTRIP-CRMP2L+ bicistronic lentivector with c-myc IRESThe bicitronic lentivector plasmid codes for renilla luciferase LucR and firefly luciferase Luc+ under the control of the cytomegalovirus (CMV) promoter, with the human c-myc IRES (363 nt-long leader from P2 promoter, Nanbru et al, J. Biol. Chem.1997) between the two reporter genes. The LucR gene used here is a modified version of LucR where all the predicted splice donor sites have been mutated. pTRIP is a Delta-U3 plasmid allowing to produce self-inactivating (SIN) lentivectorspTRIP-CRMP2L+.apepTRIP-CREL+ bicistronic lentivector with EMCV IRESThe bicitronic lentivector plasmid codes for renilla luciferase LucR and firefly luciferase Luc+ under the control of the cytomegalovirus (CMV) promoter, with the encephalomyocarditis virus (EMCV) 640 nt-long IRES (Elroy-Stein et al, PNAS 1989) between the two reporter genes. This IRES contains the T7 promoter in 5'. The LucR gene used here is a modified version of LucR where all the predicted splice donor sites have been mutated. pTRIP is a Delta-U3 plasmid allowing to produce self-inactivating (SIN) lentivectorspTRIP-CREL+.apepTRIP-CRHL+ bicistronic lentivector with no IRESThe bicitronic lentivector plasmid codes for renilla luciferase LucR and firefly luciferase Luc+ under the control of the cytomegalovirus (CMV) promoter, with a hairpin between the two reporter genes (no IRES control). The LucR gene used here is a modified version of LucR where all the predicted splice donor sites have been mutated. pTRIP is a Delta-U3 plasmid allowing to produce self-inactivating (SIN) lentivectorspTRIP-CRHL+.apeLucR-mutRenilla luciferase gene with silent point mutations on 5 AGGT splice donor sites. These mutations do not alter luciferase activity but prevent cryptic splicing events in bicistronic vectors, that could interfere with IRES activity measurement.