Journal: Journal of Advanced Research

Article Title: Tankyrase activity is essential for asymmetric division and chromosome segregation in oocyte meiosis

doi: 10.1016/j.jare.2025.07.008

Figure Lengend Snippet: TNKS regulates BubR1/MPF activity, and Securin accumulation in mouse oocyte. (A) Representative images of BubR1 signals at the kinetochore in oocytes from the control and JW55 treatment groups. Green, BubR1. Red, ACA. Blue, DNA. Bar = 5 μm. (B) The percentage of positive BubR1 signals at the kinetochore in oocytes from the control (n = 53) and JW55 (n = 50) treatment groups. ***, P < 0.001. (C) Representative images of oocytes PB1 extrusion after 12 h culture in the Control, control + Milri, control + 10 μM ETOP, JW55 + 10 μM ETOP and JW55 groups. GV oocytes were exposed to etoposide (ETOP) at concentrations of 1 μM and 10 μM for 30 min in the presence 4 μM milrinone (Milri) to prevent GVBD during this treatment. Bar = 80 μm. (D) The percentage of PB1 or large PB1 extrusion in the control (n = 94), control + Milri (n = 89), control + 1 μM ETOP (n = 98), control + 10 μM ETOP (n = 97), JW55 + 1 μ M ETOP (n = 128), JW55 + 10 μM ETOP (n = 120), and JW55 treatment (n = 114) oocytes after ETOP pretreatment. ns, P > 0.05. **, P < 0.01. ***, P < 0.001. (E) Representative images of γ-H2AX in MI stage oocyte from the control + 10 μM ETOP and JW55 + 10 μM ETOP oocytes. Red, γ-H2AX. Blue, DNA. Bar = 5 μm. (F) Relative intensity of γ-H2AX in MI stage oocyte from the control + 10 μM ETOP (n = 47) and JW55 + 10 μM ETOP (n = 39) oocytes. ***, P < 0.001. (G) Western blot of Cyclin B1 and Securin in oocytes from the control and JW55 treatment groups after 6 h of culture. (H) Western blot of pT161-CDK1 and CDC20 in oocytes from the control and JW55 treatment groups after 6 h of culture, and band intensity analysis of their protein levels in oocytes from the control and JW55 treatment groups. *, P < 0.05. **, P < 0.01. (I) Band intensity analysis of pT161-CDK1, pY15-CDK1, and Securin in oocytes from the control and JW55 treatment groups after 3 h of culture. ns, P > 0.05. *, P < 0.05. (J) Band intensity analysis of CDC25C, Cyclin B1, and pT14-CDK1 in oocytes from the control and JW55 treatment groups after 3 h of culture. *, P < 0.05. **, P < 0.01. (K) Protein-protein interaction network analysis of cell cycle-related proteins identified by mass spectrometry and MPF/Securin/CDC20/CDC25C upon STRING database. Different node color indicated core and non-core nodes, while edge thickness edge represented interaction strength. (L) Co-IP analysis with an anti-TNKS antibody. The immunoblot was probed with anti-CDC20 and anti-CDC25C antibodies. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Rabbit anti-α-Tubulin antibody (11224-1-AP), rabbit polyclonal anti-GAPDH antibody (10494-1-AP), rabbit anti-GRP78 antibody (11587-1-AP), rabbit anti-TNKS1 antibody (18030-1-AP), rabbit anti-Ran antibody (10469-1-AP), and rabbit anti-Rab11a antibody (67902-1-Ig), rabbit anti-Securin antibody (18040-1-AP), rabbit anti-CDC25C antibody (66912-1-Ig), rabbit anti-Formin2 antibody (11259-1-AP), rabbit anti-BubR1 (11504-2-AP) were from Proteintech.

Techniques: Activity Assay, Control, Western Blot, Mass Spectrometry, Co-Immunoprecipitation Assay

Journal: Drug Design, Development and Therapy

Article Title: The Effect of AZD5153 on Radiosensitivity in Pancreatic Cancer Cells Through ATM-chk1 Pathway

doi: 10.2147/DDDT.S568551

Figure Lengend Snippet: Effect of AZD5153 on protein expression in irradiated pancreatic cancer cells, n=6. ( a and b ) Western blot analysis was performed to detect the expression of BRD4, c-Myc in different groups, β-actin was used as a loading control. ( c and d ) The protein expression of p-ATM, ATM, p-chk1, chk1, p-cdc25, cdc25, p-cdc2, cdc2 in different groups was detected by western blot, β-actin was used as a loading control. Results shown are the means ± SD of 3 independent experiments with similar results for all assays. Significance was determined by Student’s t -test (* p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: The membranes were blocked with 5% skim milk for 2 h, and incubated with primary antibodies against BRD4 (Cell Signaling Technology, Cat# 13440s), c-Myc (Cell Signaling Technology, Cat# 18583s), p-ATM (Abcam, Cat# ab81292), ATM (Cell Signaling Technology, Cat# 2873s), p-cdc25C (Cell Signaling Technology, Cat# 4901s), cdc25C (Cell Signaling Technology, Cat# 4688s), p-chk1 (Cell Signaling Technology, Cat# 2348s), chk1 (Abcam, Cat# ab40866), p-cdc2 (Cell Signaling Technology, Cat# 4539s), cdc2 (Cell Signaling Technology, Cat# 9116s), γ-H2AX (Cell Signaling Technology, Cat# 9718s), cleaved PARP (Cell Signaling Technology, Cat# 5625s), Bax (Cell Signaling Technology, Cat# 14796s), β-actin (Abcam, Cat# ab6276),Vinculin (Cell Signaling Technology, Cat# 13901s) overnight at 4 °C.

Techniques: Expressing, Irradiation, Western Blot, Control

Journal: Drug Design, Development and Therapy

Article Title: The Effect of AZD5153 on Radiosensitivity in Pancreatic Cancer Cells Through ATM-chk1 Pathway

doi: 10.2147/DDDT.S568551

Figure Lengend Snippet: The effect of overexpression of chk1 on enhancing the radiosensitivity of AZD5153, n=6. ( a ) Overexpression of chk1 protein in Capan2 cells by chk1 plasmid; ( b ) Quantitative statistics of apoptosis in Capan2 cells caused by chk1 overexpression; ( c ) Representative results of the effect of chk1 overexpression on apoptosis of Capan2 cells. Results shown are the means ± SD of 3 independent experiments with similar results for all assays. Significance was determined by Student’s t -test (* p < 0.05, *** p < 0.001).

Article Snippet: The membranes were blocked with 5% skim milk for 2 h, and incubated with primary antibodies against BRD4 (Cell Signaling Technology, Cat# 13440s), c-Myc (Cell Signaling Technology, Cat# 18583s), p-ATM (Abcam, Cat# ab81292), ATM (Cell Signaling Technology, Cat# 2873s), p-cdc25C (Cell Signaling Technology, Cat# 4901s), cdc25C (Cell Signaling Technology, Cat# 4688s), p-chk1 (Cell Signaling Technology, Cat# 2348s), chk1 (Abcam, Cat# ab40866), p-cdc2 (Cell Signaling Technology, Cat# 4539s), cdc2 (Cell Signaling Technology, Cat# 9116s), γ-H2AX (Cell Signaling Technology, Cat# 9718s), cleaved PARP (Cell Signaling Technology, Cat# 5625s), Bax (Cell Signaling Technology, Cat# 14796s), β-actin (Abcam, Cat# ab6276),Vinculin (Cell Signaling Technology, Cat# 13901s) overnight at 4 °C.

Techniques: Over Expression, Plasmid Preparation