Journal: Oncotarget

Article Title: The prognostic significance of nuclear expression of PHF2 and C/EBPα in clear cell renal cell carcinoma with consideration of adipogenic metabolic evolution

doi: 10.18632/oncotarget.19949

Figure Lengend Snippet: Clinicopathologic features of patients with ccRCC and correlation between nuclear PHF2 and C/EBPα expression and clinicopathologic parameters.

Article Snippet: Each polyclonal rabbit anti-PHF2 antibody (Novus Biologicals, Littleton, CO) and polyclonal rabbit anti-C/EBPα antibody (Santa Cruz Biotechnology, Dallas, TX) was diluted 1:200.

Techniques: Expressing

Journal: Oncotarget

Article Title: The prognostic significance of nuclear expression of PHF2 and C/EBPα in clear cell renal cell carcinoma with consideration of adipogenic metabolic evolution

doi: 10.18632/oncotarget.19949

Figure Lengend Snippet: Multivariate analysis of cancer-specific and progression-free survival with PHF2 and C/EBPα nuclear expression in 344 patients with ccRCC (Cox proportional hazard model).

Article Snippet: Each polyclonal rabbit anti-PHF2 antibody (Novus Biologicals, Littleton, CO) and polyclonal rabbit anti-C/EBPα antibody (Santa Cruz Biotechnology, Dallas, TX) was diluted 1:200.

Techniques: Expressing

Journal: Scientific Reports

Article Title: HIF-1α-mediated augmentation of miRNA-18b-5p facilitates proliferation and metastasis in osteosarcoma through attenuation PHF2

doi: 10.1038/s41598-022-13660-w

Figure Lengend Snippet: miR-18b-5p modulates the expression of PHF2 through post-transcriptional regulation. ( a ) Venn diagram showed the predicted targets of miR-18b-5p from databases (miRanda, miRmap, microT and PicTar). The image was created using Online tools of Bioinformatics Evolutionary Genomics ( http://bioinformatics.psb.ugent.be/webtools/Venn/ ). ( b ) Schematic diagram showed the predicted interaction between 3′-UTR-PHF2- mRNA and miR-18b-5p. The mutated sequences of the 3′-UTR-PHF2- mRNA was displayed. ( c , d ) RT-qPCR assay showed the expression of PHF2 mRNA in MG-63/143b cells with miR-18b-5p overexpression or inhibition. ( e , f ) Western blotting assay showed the protein level of PHF2 in miR-18b-5p upregulated or downregulated MG-63 and 143b cells. ( g ) Biotin-based pulldown assay detected the enrichment of PHF2 on biotinylated miR-NC and miR-18b-5p probe with the mRNA of GAPDH acted as control. ( h , i ) The luciferase reporter assay accessed luciferase activity of miR-18b-5p upregulated or downregulated MG-63 and 143b cells transfected with luciferase reporter plasmid carrying WT-3′-UTR-PHF2 or MUT-3′-UTR-PHF2.

Article Snippet: The overexpression plasmid of PHF2 and scrambled vectors (Table ) were purchased from GeneChem (Shanghai, China).

Techniques: Expressing, Quantitative RT-PCR, Over Expression, Inhibition, Western Blot, Control, Luciferase, Reporter Assay, Activity Assay, Transfection, Plasmid Preparation

Journal: Scientific Reports

Article Title: HIF-1α-mediated augmentation of miRNA-18b-5p facilitates proliferation and metastasis in osteosarcoma through attenuation PHF2

doi: 10.1038/s41598-022-13660-w

Figure Lengend Snippet: PHF2 is responsible for miR-18b-5p-mediated cell proliferation, invasion and migration. ( a , b ) RT-qPCR and western blotting results showed the mRNA and protein expression of PHF2 in MG-63/143b cells transfected with PHF2 overexpression plasmid, miR-18b-5p mimic or negative control. ( c – e ) The proliferation, invasion and migration of MG-63/143b cells, transfected with PHF2 overexpression plasmid, miR-18b-5p mimic or negative control, was evaluated by the MTT and transwell assays. Scale bar = 100 μm.

Article Snippet: The overexpression plasmid of PHF2 and scrambled vectors (Table ) were purchased from GeneChem (Shanghai, China).

Techniques: Migration, Quantitative RT-PCR, Western Blot, Expressing, Transfection, Over Expression, Plasmid Preparation, Negative Control

Journal: Scientific Reports

Article Title: HIF-1α-mediated augmentation of miRNA-18b-5p facilitates proliferation and metastasis in osteosarcoma through attenuation PHF2

doi: 10.1038/s41598-022-13660-w

Figure Lengend Snippet: Hypoxia-driven HIF-1α is critical for upregulation of miR-18b-5p in OS. ( a ) The luciferase reporter assay accessed luciferase activity of MG-63/143b cells transfected with luciferase reporter plasmid carrying miR-18b promoter sequence or scrambled vector under normoxia and hypoxia. ( b ) RT-qPCR assay showed miR-18b-5p expression in MG-63/143b cells cultured under hypoxia for 0 h, 6 h, 12 h and 24 h. ( c ) (Up) Schematic diagram showed the predicted HRE site on the promoter of miR-18b. (Down) ChIP assay using antibodies against HIF-1α or IgG were performed to validate the binding of HIF-1α on the promoter of miR-18b. ( d ) The luciferase reporter assay accessed luciferase activity of MG-63 and 143b cells transfected with luciferase reporter plasmid carrying WT- miR-18b or MUT- miR-18b, siRNA targeting HIF-1α and negative control under normoxia or hypoxia. ( e ) Western blotting assay accessed protein level of HIF-1α in MG-63/143b cells transfected with siRNA targeting HIF-1α and negative control under normoxia or hypoxia. ( f,g ) RT-qPCR assay showed the expression of miR-18b-5p and PHF2 mRNA in MG-63/143b cells transfected with siRNA targeting HIF-1α and negative control under normoxia or hypoxia.

Article Snippet: The overexpression plasmid of PHF2 and scrambled vectors (Table ) were purchased from GeneChem (Shanghai, China).

Techniques: Luciferase, Reporter Assay, Activity Assay, Transfection, Plasmid Preparation, Sequencing, Quantitative RT-PCR, Expressing, Cell Culture, Binding Assay, Negative Control, Western Blot

Journal: Scientific Reports

Article Title: HIF-1α-mediated augmentation of miRNA-18b-5p facilitates proliferation and metastasis in osteosarcoma through attenuation PHF2

doi: 10.1038/s41598-022-13660-w

Figure Lengend Snippet: Hypoxia induced miR-18b-5p promotes OS via HIF-1α-miR-18b-5p-PHF2 axis. ( a – c ) Pearson correlation analysis showed the correlation between miR-18b-5p and HIF-1α, miR-18b-5p and PHF2, PHF2 and HIF-1α in OS tissues determined by RT-qPCR. ( d ) Schematic diagram showed the molecular mechanism of miR-18b-5p involved in OS.

Article Snippet: The overexpression plasmid of PHF2 and scrambled vectors (Table ) were purchased from GeneChem (Shanghai, China).

Techniques: Quantitative RT-PCR

Journal: Scientific Reports

Article Title: HIF-1α-mediated augmentation of miRNA-18b-5p facilitates proliferation and metastasis in osteosarcoma through attenuation PHF2

doi: 10.1038/s41598-022-13660-w

Figure Lengend Snippet: The sequence of PCR primers.

Article Snippet: The overexpression plasmid of PHF2 and scrambled vectors (Table ) were purchased from GeneChem (Shanghai, China).

Techniques: Sequencing