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KEY RESOURCES TABLE
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MedChemExpress polysome profile analysis cells
Figure 4. HIF1A-AS1 promotes HIF1a translation via recruiting RNA binding protein YB1. A, Combined immunofluorescence of the nucleolar marker protein nucleolin (yellow) and RNA-FISH <t>analysis</t> of HIF1A-AS1 (green) and HIF1a mRNA (red) in BxPC3/PANC1GEM-R <t>cells.</t> B, Schematic of affinity purification and detection of MS2-tagged HIF1A-AS1. C, GEM-R cells were transfected with the MS2 fusion plasmid system and then immunoprecipitation was performed using anti-GST antibody. The relative enrichment of HIF1a mRNA in the beads of MS2 group or HIF1A-AS1-MS2 group was assessed by qRT-PCR. D, RNA interaction profile from catRAPID suggesting that HIF1A-AS1 binds to the YB1 protein. E and F, Total cellular proteins of GEM-R cells were used in a biotin-labeled RNA pull-down assay, and YB1 binding to HIF1A-AS1 (E) or HIF1a mRNA (F) was detected via Western blot. Bio-AS1/HIF1a mRNA, transcribed and labeled HIF1A-AS1/HIF1a mRNA; No bio-AS1/HIF1a mRNA, transcribed but not labeled HIF1A-AS1/HIF1a mRNA; Ctrl RNA, transcribed and labeled antisense of HIF1A-AS1/HIF1a mRNA. G, BxPC3GEM-R cell lysates were mixed with biotinylated HIF1A-AS1 RNA or antisense (dotted line) fragments, then RNA pull-down assay was performed. (Continued on the following page.)
Polysome Profile Analysis Cells, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Journal: Cell reports

Article Title: Kaposi’s sarcoma herpesvirus activates the hypoxia response to usurp HIF2α-dependent translation initiation for replication and oncogenesis

doi: 10.1016/j.celrep.2021.110144

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Peakchart Software (polysome profiling) , Brandel , http://www.brandel.com/fractgradient.html.

Techniques: Immunohistochemistry, Virus, Recombinant, Lysis, SYBR Green Assay, Bicinchoninic Acid Protein Assay, Amplification, Reverse Transcription, Immunoprecipitation, Enzyme-linked Immunosorbent Assay, Control, Software, Real-time Polymerase Chain Reaction, Western Blot

Figure 4. HIF1A-AS1 promotes HIF1a translation via recruiting RNA binding protein YB1. A, Combined immunofluorescence of the nucleolar marker protein nucleolin (yellow) and RNA-FISH analysis of HIF1A-AS1 (green) and HIF1a mRNA (red) in BxPC3/PANC1GEM-R cells. B, Schematic of affinity purification and detection of MS2-tagged HIF1A-AS1. C, GEM-R cells were transfected with the MS2 fusion plasmid system and then immunoprecipitation was performed using anti-GST antibody. The relative enrichment of HIF1a mRNA in the beads of MS2 group or HIF1A-AS1-MS2 group was assessed by qRT-PCR. D, RNA interaction profile from catRAPID suggesting that HIF1A-AS1 binds to the YB1 protein. E and F, Total cellular proteins of GEM-R cells were used in a biotin-labeled RNA pull-down assay, and YB1 binding to HIF1A-AS1 (E) or HIF1a mRNA (F) was detected via Western blot. Bio-AS1/HIF1a mRNA, transcribed and labeled HIF1A-AS1/HIF1a mRNA; No bio-AS1/HIF1a mRNA, transcribed but not labeled HIF1A-AS1/HIF1a mRNA; Ctrl RNA, transcribed and labeled antisense of HIF1A-AS1/HIF1a mRNA. G, BxPC3GEM-R cell lysates were mixed with biotinylated HIF1A-AS1 RNA or antisense (dotted line) fragments, then RNA pull-down assay was performed. (Continued on the following page.)

Journal: Cancer Research

Article Title: LncRNA HIF1A-AS1 Promotes Gemcitabine Resistance of Pancreatic Cancer by Enhancing Glycolysis through Modulating the AKT/YB1/HIF1α Pathway

doi: 10.1158/0008-5472.can-21-0281

Figure Lengend Snippet: Figure 4. HIF1A-AS1 promotes HIF1a translation via recruiting RNA binding protein YB1. A, Combined immunofluorescence of the nucleolar marker protein nucleolin (yellow) and RNA-FISH analysis of HIF1A-AS1 (green) and HIF1a mRNA (red) in BxPC3/PANC1GEM-R cells. B, Schematic of affinity purification and detection of MS2-tagged HIF1A-AS1. C, GEM-R cells were transfected with the MS2 fusion plasmid system and then immunoprecipitation was performed using anti-GST antibody. The relative enrichment of HIF1a mRNA in the beads of MS2 group or HIF1A-AS1-MS2 group was assessed by qRT-PCR. D, RNA interaction profile from catRAPID suggesting that HIF1A-AS1 binds to the YB1 protein. E and F, Total cellular proteins of GEM-R cells were used in a biotin-labeled RNA pull-down assay, and YB1 binding to HIF1A-AS1 (E) or HIF1a mRNA (F) was detected via Western blot. Bio-AS1/HIF1a mRNA, transcribed and labeled HIF1A-AS1/HIF1a mRNA; No bio-AS1/HIF1a mRNA, transcribed but not labeled HIF1A-AS1/HIF1a mRNA; Ctrl RNA, transcribed and labeled antisense of HIF1A-AS1/HIF1a mRNA. G, BxPC3GEM-R cell lysates were mixed with biotinylated HIF1A-AS1 RNA or antisense (dotted line) fragments, then RNA pull-down assay was performed. (Continued on the following page.)

Article Snippet: Polysome profile analysis Cells were incubated with 100 mg/mL cycloheximide (MedChem- Express) at 37 C for 30 minutes, and then were pelleted and lysed on ice with polysome lysis buffer.

Techniques: RNA Binding Assay, Marker, Transfection, Plasmid Preparation, Immunoprecipitation, Quantitative RT-PCR, Labeling, Pull Down Assay, Binding Assay, Western Blot