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Journal: bioRxiv
Article Title: Identification of transcriptome-wide cobalt chloride-induced hypoxia-responsive long noncoding RNAs regulated by cytoplasmic mRNA capping enzyme
doi: 10.1101/2024.11.21.624636
Figure Lengend Snippet: (A) The selected lncRNAs possess multiple HIF1α binding motifs (R/CGTG) within their promoter regions, as illustrated in the schematic. (B) Western blot shows the addition of CoCl 2 led to elevated expression of HIFα which was reduced after PX-478 treatment. (C) The RT–qPCR analysis of cytoplasmic RNA from the above treatment groups showed that treating U2OS cells with the HIF1α-specific inhibitor PX-478 under hypoxic conditions led to a significant decrease in the expression of LUCAT1-211 and MSC-AS1-212. CA9 serves as a known hypoxia-induced gene regulated by HIF1α. Values are expressed as mean ± standard deviation (SD) from three biological replicates. Statistical significance is denoted as follows: ns indicates non-significant; *P < 0.05; **P < 0.005; ***P < 0.0005; ****P < 0.0001, with n ≥ 3.
Article Snippet: The primary antibodies used included: 1:1000 rabbit anti-HIF1α (Cell Signaling Technology, cat no# 36169), 1:1000 mouse anti-β actin (Santa Cruz Biotechnology, cat no# SC-517582), 1:1000 rabbit anti-CE (Novus Biologicals, cat no# NPB1-49973), 1:2000 mouse anti-GAPDH (Novus Biologicals, cat no# 2D4A7), 1:2000
Techniques: Binding Assay, Western Blot, Expressing, Quantitative RT-PCR, Standard Deviation