Rabbit anti hif 1α antibody

RPMI1640, fetal bovine serum and trypsase all were bought from Sigma Co.Ltd (USA). RNeasy Mini Kit were purchased from Hilden Co.Ltd (Germany). Arraystar Human circRNAs chip were purchased from Rockville Co. Ltd(USA). TRIzol reagent, MTT, DMSO, crystal violet, PCR reverse transcription kit were bought from Invitrogen Co.Ltd (USA). Annexin V/PI cell apoptosis kits were purchased from Sigma Co.Ltd (USA). Rabbit anti-HIF-1α antibody (1:1000 dilution, catalog number: ab203848), and Goat Anti-Rabbit IgG H&L (HRP) (1:1000, catalog number: ab205718) were purchased from Abcam Co.Ltd (USA).

Mouse anti-E-cadherin and anti-vimentin antibodies were purchased from BD Technologies (La Jolla, CA, USA). Rabbit anti-ZO1 antibody was purchased from Invitrogen (Carlsbad, CA, USA). Rabbit anti-Snail antibodies, DMOG and LY294002 were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). Antibodies against Slug, GSK3β, p-GSK3β (Ser9), Akt, p-Akt (Ser473), STAT3, and p-STAT3 (Thr705) were purchased from Cell Signaling Technology (Beverly, MA, USA). Rabbit anti-HIF-1α antibody was purchased from Abcam (Cambridge, MA, USA), and mouse anti-beta-actin antibody was obtained from Proteintech Group (Chicago, IL, USA). Goat anti-mouse (W4021) and goat anti-rabbit (W4011) peroxidase-conjugated secondary antibodies were obtained from Pierce Biotechnology (Rockford, IL, USA). The following reagents were used: anti-IL-11 neutralizing antibody (LifeSpan Technology, Newton, MA, USA), recombinant human IL-11 (rhIL-11) (Prospecbio, Rehovot, Israel), and Akt inhibitor V (Millipore, Billerica, MA, USA). All other reagents, unless otherwise specified, were purchased from Sigma-Aldrich (St. Louis, MO, USA).

The ChIP assay was performed as described previously⁶² . ChIP was performed on MDA-MB-231 cells exposed to normoxia (21% O₂) or hypoxia (0.5% O₂) for 24 h using rabbit anti-HIF1α antibody (Abcam). Normal rabbit IgG (Santa Cruz Biotechnology) was used as a nonspecific IgG control. Cells were formalin fixed, lysed, and sonicated using a Bioruptor Plus Sonication System (Diagenode Inc.) set at high power. Sonicated chromatin was incubated overnight with antibodies followed by precipitation with protein A/G Dynabeads (Invitrogen). Five percent of the sample from each immunoprecipitation was reserved for input control. Alternatively, relative enrichment was measured by quantitative RT-PCR using a titration of pooled input samples as a standard curve and normalized to input after subtraction of IgG signal. Enrichments are presented as fold change relative to normoxia or IgG control. Each primer sequence is indicated in Supplementary Data 5.