Journal: Cell & bioscience

Article Title: SETD8, a frequently mutated gene in cervical cancer, enhances cisplatin sensitivity by impairing DNA repair.

doi: 10.1186/s13578-023-01054-y

Figure Lengend Snippet: Fig. 3 SETD8 inhibition affects 53BP1 accumulation by reducing the methylation level of H4K20. (A) Western blot analysis of SETD8 and H4K20me1/ H4K20me2 levels. Cells transfected with two different SETD8 siRNAs were treated with cisplatin for 24 h compared to siNC. (B) Immunofluorescence for H4K20me1 and 53BP1 post cisplatin in SiHa and CaSki cells, Scale bar, 10 μm. (C) Immunofluorescence for H4K20me2 and 53BP1 post cisplatin in SiHa and CaSki cells, Scale bar, 10 μm. (D-E) Plot profiles show fluorescence intensity along an oblique line quantified by ImageJ. (F-G) Pearson’s Coefficient and Overlap Coefficient calculated with JACoP by ImageJ. Pearson’s Coefficient or Overlap Coefficient > 0.5 indicates co-localization

Article Snippet: Membranes were blocked with 5% milk-TBST and incubated with primary antibodies against the following proteins: SETD8 (Proteintech, 14063-1-AP, 1:500), H4K20me1 (Abclonal, A2370, 1:1000), H4K20me2 (Abclonal, A2371, 1:1000), GAPDH (Abclonal, AC001, 1:20000), 53BP1 (Abclonal, A5757, 1:500), and γ-H2AX (Abclonal, AP0245, 1:1000).

Techniques: Inhibition, Methylation, Western Blot, Transfection, Immunofluorescence, Fluorescence

Journal: Cell & bioscience

Article Title: SETD8, a frequently mutated gene in cervical cancer, enhances cisplatin sensitivity by impairing DNA repair.

doi: 10.1186/s13578-023-01054-y

Figure Lengend Snippet: Fig. 4 Reduction of 53BP1 binding to DNA breaks induced by SETD8 downregulation leads to cisplatin sensitivity by inhibiting NHEJ. (A-B) Accumulation of γ-H2AX and 53BP1 following SETD8 silencing in cisplatin-treated SiHa cells for 24 h and quantification of γ-H2AX and 53BP1 foci. (C-D) Accumulation of γ-H2AX and 53BP1 following SETD8 silencing in cisplatin-treated CaSki cells for 24 h and quantification of γ-H2AX and 53BP1 foci. (E) Schematic diagram for DSB Repair Reporter. (F) NHEJ efficiency detected by DSB Repair Reporter. Cells with or without SETD8 siRNA were treated with cisplatin for 24 h. p values were determined by two-tailed Student’s t test (ns: not significant; *: 0.01 ≤ p < 0.05; **: 0.001 ≤ p < 0.01; ***: 0.0001 ≤ p < 0.001; ****: p < 0.0001)

Article Snippet: Membranes were blocked with 5% milk-TBST and incubated with primary antibodies against the following proteins: SETD8 (Proteintech, 14063-1-AP, 1:500), H4K20me1 (Abclonal, A2370, 1:1000), H4K20me2 (Abclonal, A2371, 1:1000), GAPDH (Abclonal, AC001, 1:20000), 53BP1 (Abclonal, A5757, 1:500), and γ-H2AX (Abclonal, AP0245, 1:1000).

Techniques: Binding Assay, Two Tailed Test

Journal: Cell & bioscience

Article Title: SETD8, a frequently mutated gene in cervical cancer, enhances cisplatin sensitivity by impairing DNA repair.

doi: 10.1186/s13578-023-01054-y

Figure Lengend Snippet: Fig. 7 Schematic diagram of SETD8 inhibition sensitizes cervical cancer cells to cisplatin treatment. Downregulation of SETD8 significantly reduces the mono-methylation level of H4K20 (H4K20me1) and double-methylation level of H4K20 (H4K20me2) in cervical cancer and further reduces the recruit ment of the DNA damage repair factor 53BP1, inhibits the non-homologous end joining repair pathway (NHEJ), resulting in increased cisplatin-induced DNA damage

Article Snippet: Membranes were blocked with 5% milk-TBST and incubated with primary antibodies against the following proteins: SETD8 (Proteintech, 14063-1-AP, 1:500), H4K20me1 (Abclonal, A2370, 1:1000), H4K20me2 (Abclonal, A2371, 1:1000), GAPDH (Abclonal, AC001, 1:20000), 53BP1 (Abclonal, A5757, 1:500), and γ-H2AX (Abclonal, AP0245, 1:1000).

Techniques: Inhibition, Methylation, Non-Homologous End Joining

Journal: Cancers

Article Title: Testicular Germ Cell Tumors Acquire Cisplatin Resistance by Rebalancing the Usage of DNA Repair Pathways

doi: 10.3390/cancers13040787

Figure Lengend Snippet: List of the antibodies, commercial sources and dilutions used in this study.

Article Snippet: TP53BP1 , OriGene , TA309918 , 1:1000.

Techniques:

Journal: Cancers

Article Title: Testicular Germ Cell Tumors Acquire Cisplatin Resistance by Rebalancing the Usage of DNA Repair Pathways

doi: 10.3390/cancers13040787

Figure Lengend Snippet: List of the antibodies, commercial sources and dilutions used in this study.

Article Snippet: Other primary antibodies used were as follows: anti-H2AX Ser139 (1:250; Cell Signaling 978), anti-RPA1 (1:250; Santa Cruz Biotechnology B6), anti-RAD51 (1:250 H-92; Santa Cruz Biotechnology), anti-BRCA1 (1:100 D9; Santa Cruz Biotechnology), anti-FANCD2 (1:250; Novus Biologicals, Centennial, CO, USA; NB100-182) and anti-53BP1 (1:250; OriGene, Rockville, MD, USA; ta309918).

Techniques: