Journal: Cancer Medicine

Article Title: PHD finger protein 5A promoted lung adenocarcinoma progression via alternative splicing

doi: 10.1002/cam4.2115

Figure Lengend Snippet: PHF5A upregulation is associated with poor patient survival in The Cancer Genome Atlas (TCGA) data sets. A, PHF5A expression was significantly increased in NSCLC tumor tissues (n = 1016) compared with normal tissues (n = 110). B and C, PHF5A expression was significantly increased in lung adenocarcinoma (LUAD) (B, n = 58) and lung squamous cell carcinoma (LUSC) (C, n = 51) tumor tissues compared with paired adjacent normal tissues. D and E, A high mRNA level of PHF5A was significantly associated with shorter OS in LUAD patients (D) but not LUSC patients (E). * P < 0.05; *** P < 0.001

Article Snippet: For overexpression, the open reading frame (ORF) of PHF5A with C terminal Flag and His tag was compounded by Vigene (Shandong, China) and ligated into the pEnter vector; the empty vector was used as negative control.

Techniques: Expressing

Journal: Cancer Medicine

Article Title: PHD finger protein 5A promoted lung adenocarcinoma progression via alternative splicing

doi: 10.1002/cam4.2115

Figure Lengend Snippet: The expression of PHF5A is significantly upregulated in both lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC) patient samples. A and B, The percentage of PHF5A‐low and PHF5A‐high staining in tumor and paired normal tissues in LUAD (A, n = 60) and LUSC (B, n = 66) patients. C, The number and percentage of patients with higher, equal or lower PHF5A staining in LUAD and LUSC tumor tissues compared with paired normal tissues. T: tumor tissue; N: paired normal tissue. D, Representative IHC images of PHF5A staining in NSCLC tumor tissues and paired normal tissues. E and F, Kaplan‐Meier plots based on the expression level of PHF5A measured by IHC in 58 LUAD (E) and 63 LUSC (F) patients

Article Snippet: For overexpression, the open reading frame (ORF) of PHF5A with C terminal Flag and His tag was compounded by Vigene (Shandong, China) and ligated into the pEnter vector; the empty vector was used as negative control.

Techniques: Expressing, Staining

Journal: Cancer Medicine

Article Title: PHD finger protein 5A promoted lung adenocarcinoma progression via alternative splicing

doi: 10.1002/cam4.2115

Figure Lengend Snippet: Downregulation of PHF5A inhibited lung adenocarcinoma (LUAD) cell proliferation, invasion and migration. A, The knockdown efficiency of PHF5A was examined by qRT‐PCR and western blot. GAPDH was used as a control. B, The proliferation ability of H1299 and A549 cell lines transfected with siRNAs was measured by CCK8 assay. C, Clone formation assays in PHF5A knockdown H1299 and A549 cell lines and control cells. D, Cell cycle analysis in PHF5A knockdown H1299 and A549 cell lines and control cells. E, Cisplatin–induced apoptosis in PHF5A knockdown H1299 and A549 cell lines and control cells. F, The migration and invasion ability in PHF5A knockdown H1299 and A549 cell lines and control cells. Data are expressed as the means ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Student's t test

Article Snippet: For overexpression, the open reading frame (ORF) of PHF5A with C terminal Flag and His tag was compounded by Vigene (Shandong, China) and ligated into the pEnter vector; the empty vector was used as negative control.

Techniques: Migration, Knockdown, Quantitative RT-PCR, Western Blot, Control, Transfection, CCK-8 Assay, Cell Cycle Assay

Journal: Cancer Medicine

Article Title: PHD finger protein 5A promoted lung adenocarcinoma progression via alternative splicing

doi: 10.1002/cam4.2115

Figure Lengend Snippet: Overexpression of PHF5A promoted lung adenocarcinoma (LUAD) cell proliferation, invasion and migration. A, The overexpression efficiency of PHF5A was examined by qRT‐PCR and western blot. GAPDH was used as the control. B, The proliferation ability of H1299 and A549 cell lines transfected with PHF5A and control vectors was measured by CCK8 assay. C, Clone formation assays in PHF5A overexpressed H1299 and A549 cell lines and control cells. D, Cell cycle analysis in PHF5A–overexpressed H1299 and A549 cell lines and control cells. E, Cisplatin–induced apoptosis in PHF5A–overexpressed H1299 and A549 cell lines and control cells. F, The migration and invasion ability in PHF5A–overexpressed H1299 and A549 cell lines and control cells. Data are expressed as the means ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Student's t test

Article Snippet: For overexpression, the open reading frame (ORF) of PHF5A with C terminal Flag and His tag was compounded by Vigene (Shandong, China) and ligated into the pEnter vector; the empty vector was used as negative control.

Techniques: Over Expression, Migration, Quantitative RT-PCR, Western Blot, Control, Transfection, CCK-8 Assay, Cell Cycle Assay

Journal: Cancer Medicine

Article Title: PHD finger protein 5A promoted lung adenocarcinoma progression via alternative splicing

doi: 10.1002/cam4.2115

Figure Lengend Snippet: Knockdown of PHF5A–induced genome–wide alternative splicing (AS) events. A, The number of significantly differentially spliced events in five AS types. B, GO enrichment analysis in differentially spliced genes. C, KEGG pathway enrichment analysis in differentially spliced genes. D, Validation of AS events in cell cycle–related genes using RT‐PCR. E, Validation of AS events in apoptosis–related genes using RT‐PCR. F, Validation of AS events in other differentially spliced genes using RT‐PCR

Article Snippet: For overexpression, the open reading frame (ORF) of PHF5A with C terminal Flag and His tag was compounded by Vigene (Shandong, China) and ligated into the pEnter vector; the empty vector was used as negative control.

Techniques: Knockdown, Genome Wide, Alternative Splicing, Biomarker Discovery, Reverse Transcription Polymerase Chain Reaction

Journal: Cancer Medicine

Article Title: PHD finger protein 5A promoted lung adenocarcinoma progression via alternative splicing

doi: 10.1002/cam4.2115

Figure Lengend Snippet: Inhibitor of PHF5A inhibited lung adenocarcinoma (LUAD) cell proliferation and induced similar alternative splicing (AS) events. A, The IC50 curve of pladienolide in H1299 and A549 cell lines. B and C, CCK8 assays in H1299 (B) and A549 (C) cell lines treated with different concentrations of pladienolide. D and E, Validation of AS events in partial genes in H1299 (D) and A549 (E) cell lines treated with different concentrations of pladienolide. **** P < 0.0001

Article Snippet: For overexpression, the open reading frame (ORF) of PHF5A with C terminal Flag and His tag was compounded by Vigene (Shandong, China) and ligated into the pEnter vector; the empty vector was used as negative control.

Techniques: Alternative Splicing, Biomarker Discovery