Journal: Oncology Letters

Article Title: Association between APOBEC3s and HPV16 E2 gene hypermutation in Uygur females with cervical cancer

doi: 10.3892/ol.2020.11697

Figure Lengend Snippet: PCR primer sequences.

Article Snippet: After blocking with 5% BSA at room temperature for 1 h, the membrane was incubated with anti-APOBEC3G (1:500; bs-15407R; BIOSS), anti-APOBEC3F (1:500; ab227962; Abcam), anti-APOBEC3C (1:500; bs-12495R; BIOSS) and anti-GAPDH (1:5,000; bs-50549R; BIOSS) primary antibodies at 4°C for 16 h. Subsequently, the membranes were incubated with the secondary antibody of goat anti-rabbit HRP conjugated IgG (1:10,000; ZB-2301; OriGene Technologies, Inc.) at 37°C for 1 h in the dark.

Techniques:

Journal: Oncology Letters

Article Title: Association between APOBEC3s and HPV16 E2 gene hypermutation in Uygur females with cervical cancer

doi: 10.3892/ol.2020.11697

Figure Lengend Snippet: Expression levels of APBOEC3s mRNA in cervical lesion samples of the cervicitis, CIN I–III and cervical cancer groups. The relative mRNA expression levels of (A) A3C, (B) A3F and (C) A3G in the cervical lesions of different grades were detected using reverse transcription-quantitative PCR. *P<0.05, **P<0.01. APBOEC3s, apolipoprotein B mRNA-editing enzyme catalytic polypeptides; CIN I–III, cervical intraepithelial neoplasia I, II and III.

Article Snippet: After blocking with 5% BSA at room temperature for 1 h, the membrane was incubated with anti-APOBEC3G (1:500; bs-15407R; BIOSS), anti-APOBEC3F (1:500; ab227962; Abcam), anti-APOBEC3C (1:500; bs-12495R; BIOSS) and anti-GAPDH (1:5,000; bs-50549R; BIOSS) primary antibodies at 4°C for 16 h. Subsequently, the membranes were incubated with the secondary antibody of goat anti-rabbit HRP conjugated IgG (1:10,000; ZB-2301; OriGene Technologies, Inc.) at 37°C for 1 h in the dark.

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: Oncology Letters

Article Title: Association between APOBEC3s and HPV16 E2 gene hypermutation in Uygur females with cervical cancer

doi: 10.3892/ol.2020.11697

Figure Lengend Snippet: Protein expression levels of APBOEC3 in cervical cancer of different grades. (A) Protein expression levels of A3C, A3F and A3G in cervical lesion of different grades detected using western blot analysis. Relative protein expression levels of (B) A3C, (C) A3F and (D) A3G. **P<0.01. APBOEC3s, Apolipoprotein B mRNA-editing enzyme catalytic polypeptides; CIN I–III, cervical intraepithelial neoplasia I, II and III.

Article Snippet: After blocking with 5% BSA at room temperature for 1 h, the membrane was incubated with anti-APOBEC3G (1:500; bs-15407R; BIOSS), anti-APOBEC3F (1:500; ab227962; Abcam), anti-APOBEC3C (1:500; bs-12495R; BIOSS) and anti-GAPDH (1:5,000; bs-50549R; BIOSS) primary antibodies at 4°C for 16 h. Subsequently, the membranes were incubated with the secondary antibody of goat anti-rabbit HRP conjugated IgG (1:10,000; ZB-2301; OriGene Technologies, Inc.) at 37°C for 1 h in the dark.

Techniques: Expressing, Western Blot

Journal: Oncology Letters

Article Title: Association between APOBEC3s and HPV16 E2 gene hypermutation in Uygur females with cervical cancer

doi: 10.3892/ol.2020.11697

Figure Lengend Snippet: HPV16 E2 gene expression and hypermutation in SiHa cells transfected with APOBEC3s lentiviral vectors. (A) GFP-labeled A3C, A3F and A3G lentiviruses were transfected into SiHa cells and the fluorescence intensity was observed using an inverted fluorescence microscope to determine the transfection efficiency. Magnification, ×200. The mRNA and protein expression levels of HPV16 E2 in A3 lentivirus-transfected SiHa cells were confirmed using reverse transcription-quantitative PCR and western blotting. (B) The HPV16 E2 genome hypermutation following lentiviral transfection was detected using the differential DNA Denaturation PCR. **P<0.01 vs. blank control or scrambled siRNA control. HPV, human papilloma virus; APBOEC3s, Apolipoprotein B mRNA-editing enzyme catalytic polypeptides; siRNA, small interfering RNA; NC, negative control; A3C, APOBEC3C; A3F, APOBEC3F; A3G, APOBEC3G; GFP, green fluorescent protein; C, cytosine; G, guanine; A, adenine; T, thymine.

Article Snippet: After blocking with 5% BSA at room temperature for 1 h, the membrane was incubated with anti-APOBEC3G (1:500; bs-15407R; BIOSS), anti-APOBEC3F (1:500; ab227962; Abcam), anti-APOBEC3C (1:500; bs-12495R; BIOSS) and anti-GAPDH (1:5,000; bs-50549R; BIOSS) primary antibodies at 4°C for 16 h. Subsequently, the membranes were incubated with the secondary antibody of goat anti-rabbit HRP conjugated IgG (1:10,000; ZB-2301; OriGene Technologies, Inc.) at 37°C for 1 h in the dark.

Techniques: Expressing, Transfection, Labeling, Fluorescence, Microscopy, Real-time Polymerase Chain Reaction, Western Blot, Small Interfering RNA, Negative Control