Journal: Molecular Medicine Reports

Article Title: CDK1-induced regulation of p53 phosphorylation at Ser315 mediates cell cycle arrest and apoptosis of macrophages infected with clinical isolates of Mycobacterium tuberculosis

doi: 10.3892/mmr.2025.13754

Figure Lengend Snippet: Inhibition of p53 (Ser315) phosphorylation attenuates macrophage G 2 /M cycle block and apoptosis caused by MTB infection. (A) Protein expression levels of total p53 and p-p53 (Ser315) after MTB infection were determined by western blotting. (B) Statistical analysis of total p53 and p-p53 (Ser315) expression detected by western blotting after MTB infection. (C) Protein expression levels of total p53 and p-p53 (Ser315) were determined by western blotting before and after the addition of Pft-α. (D) Statistical analysis of total p53 and p-p53 (Ser315) expression determined by western blotting before and after the addition of Pft-α. Cell cycle profiles of macrophages in the (E) V group, (F) L3 group, (G) L4 group and (H) L5 group when Pift-α was not added. Cell cycle profiles of macrophages in the (I) V group, (J) L3 group, (K) L4 group and (L) L5 group after the addition of Pft-α. Apoptosis of macrophages in the (M) V group, (N) L3 group, (O) L4 group and (P) L5 group when Pft-α was not added. Apoptosis of macrophages in the (Q), V group, (R) L3 group, (S) L4 group and (T) L5 group after the addition of Pft-α. (U) G 2 /M cell cycle ratio and (V) apoptosis rate of each group before and after the addition of Pft-α. (W) Protein expression levels of BCL2, BAX, caspase 3 and cleaved-caspase 3 were determined by western blotting before and after the addition of Pft-α. (X) Statistical analysis of apoptosis-related protein expression detected by western blotting before and after the addition of Pft-α. Data are presented as the mean ± SEM. *P<0.05, **P<0.01, ***P<0.001. A.U., arbitrary units; L group, MTB from Xinjang clinical isolate-infected group; MTB, Mycobacterium tuberculosis ; n.s., not significant; p-, phosphorylated; Pft-α, pifithrin-α; V group, H37Rv standard strain-infected group.

Article Snippet: For IP, 5 μg rabbit IgG antibody (cat. no. HY- P80879 ; MedChemExpress), 5 μg rabbit CDK1 antibody (cat. no. HY- P80611 ; MedChemExpress) and 5 μg rabbit p53 antibody (cat. no. HY- P86169 ; MedChemExpress) were mixed with 500 μg protein solution, and incubated overnight at 4°C.

Techniques: Inhibition, Phospho-proteomics, Blocking Assay, Infection, Expressing, Western Blot

Journal: Molecular Medicine Reports

Article Title: CDK1-induced regulation of p53 phosphorylation at Ser315 mediates cell cycle arrest and apoptosis of macrophages infected with clinical isolates of Mycobacterium tuberculosis

doi: 10.3892/mmr.2025.13754

Figure Lengend Snippet: Secretion of cytokines and NO before and after inhibition of CDK1 expression or inhibition of p53 (Ser315) phosphorylation. Secretion of (A) IL-6, (B) TNF-α, (C) IL-1β, (D) IL-10, (E) IL-12 and (F) NO before and after inhibition of CDK1 expression or inhibition of p53 (Ser315) phosphorylation. Data are presented as the mean ± SEM. ***P<0.001. MTB, Mycobacterium tuberculosis ; NO, nitric oxide; ns, not significant; Pft-α, pifithrin-α; XJMTB, MTB from Xinjang.

Article Snippet: For IP, 5 μg rabbit IgG antibody (cat. no. HY- P80879 ; MedChemExpress), 5 μg rabbit CDK1 antibody (cat. no. HY- P80611 ; MedChemExpress) and 5 μg rabbit p53 antibody (cat. no. HY- P86169 ; MedChemExpress) were mixed with 500 μg protein solution, and incubated overnight at 4°C.

Techniques: Inhibition, Expressing, Phospho-proteomics

Journal: Molecular Medicine Reports

Article Title: CDK1-induced regulation of p53 phosphorylation at Ser315 mediates cell cycle arrest and apoptosis of macrophages infected with clinical isolates of Mycobacterium tuberculosis

doi: 10.3892/mmr.2025.13754

Figure Lengend Snippet: CDK1 mediates G 2 /M cell cycle arrest and apoptosis caused by infection with XJMTB through regulation of p53 (Ser315) phosphorylation. (A) Phosphorylation of p53 (Ser315) after inhibition of CDK1. (B) Phosphorylation of CDK1 (Thr161) and of p53 (Ser315) following treatment with TC11. (C) Expression of CDK1 after inhibition of p53 (Ser315) phosphorylation. (D) Statistical analysis of total p53 and p-p53 (Ser315) expression determined by western blotting before and after the addition of Ro3306. (E) Statistical analysis of total p53, p-p53 (Ser315), total CDK1 and p-CDK1 (Thr161) expression determined by western blotting before and after the addition of TC11. (F) Statistical analysis of total p53, p-p53 (Ser315) and total CDK1 expressiondetermined by western blotting before and after the addition of Pft-α. Cell cycle progression of XJMTB-infected macrophages in the (G) TC11, (H) Pft-α and (I) TC11 + Pft-α groups. (J) G 2 /M cell cycle ratio in the various groups. Apoptosis of XJMTB-infected macrophages in the (K) TC11, (L) Pft-α and (M) TC11 + Pft-α groups. (N) Apoptosis rate in the various groups. (O and P) Protein expression levels of BCL2, BAX, caspase 3 and cleaved-caspase 3 were determined by western blotting in the TC11, Pft-α and TC11 + Pft-α groups. (Q) CFU analysis was performed to determine the survival rate of XJMTB in macrophages in the TC11, Pft-α and TC11 + Pft-α groups. (R) Co-IP results of CDK1 and p53. Data are presented as the mean ± SEM. *P<0.05, **P<0.01, ***P<0.001. A.U., arbitrary units; CDK1, cyclin-dependent kinase; CFU, colony-forming unit; IB, immunoblotting; IgG, immunoglobulin G; IP, immunoprecipitation; L group, XJMTB clinical isolate-infected group; MTB, Mycobacterium tuberculosis ; n.s., not significant; p-, phosphorylated; Pft-α, pifithrin-α; XJMTB, MTB from Xinjang.

Article Snippet: For IP, 5 μg rabbit IgG antibody (cat. no. HY- P80879 ; MedChemExpress), 5 μg rabbit CDK1 antibody (cat. no. HY- P80611 ; MedChemExpress) and 5 μg rabbit p53 antibody (cat. no. HY- P86169 ; MedChemExpress) were mixed with 500 μg protein solution, and incubated overnight at 4°C.

Techniques: Infection, Phospho-proteomics, Inhibition, Expressing, Western Blot, Co-Immunoprecipitation Assay, Immunoprecipitation

Journal: Journal of Virology

Article Title: p53-mediated regulation of electron transport chain and nucleotide synthesis during Newcastle disease virus infection

doi: 10.1128/jvi.01576-25

Figure Lengend Snippet: p53-mediated ETC stabilization promotes NDV replication ( A and B ) A549 cells were transfected with siRNA to knock down TP53. Twenty-four hours after transfection, cells were infected with NDV at an MOI of 1 for 6 hpi. OCR was measured using the Agilent Seahorse XFe96 extracellular flux analyzer ( A ). Basal/maximal respiration and ATP production were analyzed using GraphPad software ( B ). ( C and D ) A549 cells were transfected with siRNA to knock down TP53 and negative control siRNA (siNC). Twenty-four hours after transfection, cells were infected with NDV at an MOI of 1 for 18 hpi. Intracellular ROS levels were quantified via flow cytometry using DCFH-DA staining ( C ), with fluorescence intensity analyzed by FlowJo software ( D ). ( E and F ) H1299 cells were transfected with HA-p53. Twenty-four hours after transfection, cells were infected with NDV at an MOI of 1 for 18 hpi. Intracellular ROS levels were quantified via flow cytometry using DCFH-DA staining ( E ), with fluorescence intensity analyzed by FlowJo software ( F ). ( G–I ) A549 cells were transfected with siRNA to knock down TP53 and negative control siRNA (siNC). Twenty-four hours after transfection, cells were infected with NDV at an MOI of 1 for 12 hpi and were treated with varying concentrations of ETC inhibitors (rotenone, antimycin A, and FCCP) during this period. NC represents cells treated with DMSO. Western blotting was employed to detect the protein levels of NDV-NP, TP53, and β-actin ( G ). Cell supernatant was collected and the viral titer was determined by plaque assay ( H and I ). ( J–L ) H1299 cells were transfected with HA-p53. Twenty-four hours after transfection, cells were infected with NDV at an MOI of 1 for 12 hpi and were treated with varying concentrations of ETC inhibitors (rotenone, antimycin A, and FCCP) during this period. NC represents cells treated with DMSO. Western blotting was employed to detect the protein levels of NDV-NP, HA, and β-actin ( J ). Cell supernatant was collected and the viral titer was determined by plaque assay ( K and L ). Data are presented as means from three independent experiments. ns, no significance, * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Mouse monoclonal anti-β-actin (66009-1-Ig) and rabbit polyclonal anti-p53 (10442-1-AP) were purchased from Proteintech.

Techniques: Transfection, Knockdown, Infection, Software, Negative Control, Flow Cytometry, Staining, Fluorescence, Western Blot, Plaque Assay

Journal: Journal of Virology

Article Title: p53-mediated regulation of electron transport chain and nucleotide synthesis during Newcastle disease virus infection

doi: 10.1128/jvi.01576-25

Figure Lengend Snippet: Model of NDV-induced p53 stabilization supporting ETC activity and nucleotide synthesis for viral replication. In the presence of p53, cells adapt to virus-induced metabolic stress and maintain nucleotide pools, supporting both cell survival and viral replication. Without p53, cells become reliant on intact ETC; ETC disruption quickly depletes nucleotides and impairs viral replication. Thus, while p53 preserves cellular stability, viruses may exploit this environment to enhance their own replication.

Article Snippet: Mouse monoclonal anti-β-actin (66009-1-Ig) and rabbit polyclonal anti-p53 (10442-1-AP) were purchased from Proteintech.

Techniques: Activity Assay, Virus, Disruption

Journal: Molecular Medicine Reports

Article Title: Pan-cancer analysis of the carcinogenic role of WSB2 in human tumors

doi: 10.3892/mmr.2025.13625

Figure Lengend Snippet: WSB2 promotes the proliferation and migration of breast cancer cells. (A) Reverse transcription-quantitative PCR and (B) western blotting were used to detect the effect of WSB2 overexpression in MDA-MB-231 and MCF-7 cells. (C) Viability of MDA-MB-231 and MCF-7 cells was evaluated using a CCK-8 assay. (D) Representative images and (E) the quantification of the number of MDA-MB-231 and MCF-7 cells in the S phase evaluated using an 5 ethynyl-2′-deoxyuridine assay. (F) Representative images of colony formation ability of MDA-MB-231 and MCF-7cells. (G) Representative images and (H) quantification of wound healing and (I) colony formation ability of MDA-MB-231 and MCF-7cells. (J) Transwell assays to determine the migration ability of MDA-MB-231and MCF-7cells. (K) ERK1/2, p53 and PCNA and (L) Snail, E-cadherin and Vimentin protein levels in MDA-MB-231 and MCF-7 cells were detected using western blotting. *P<0.05, **P<0.01, ***P<0.001. WSB2, WD repeat and SOCS box containing 2.

Article Snippet: Standard technique was used to carry out western blotting ( ) and the following specific primary antibodies were used: Anti-WSB2 (cat. no. ab127176; Abcam, 1:2,000), anti-GADPH (cat. no. CSB-MA000071Mom; Cusabio Technology, LLC, 1:10,000), anti-β-Actin (cat. no. AC026; ABclonal Biotech Co., Ltd. 1:10,000 dilution), anti-HA-tag (cat. no. B1021; Suzhou Botelon Immunotechnology Co., Ltd. 1:5,000), anti-E-cadherin (cat. no. BD-PT1454; Suzhou Botelon Immunotechnology Co., Ltd.; 1:500 dilution), anti-proliferating cell nuclear antigen (PCNA; cat. no. D220014-0025; Sangon Biotech Co., Ltd. 1:1,000 dilution), anti-Vimentin (cat. no. BD-PB4686; Suzhou Botelon Immunotechnology Co., Ltd. 1:500), anti-p53 antibody (p53) (cat. no. CSB-PA07889A0Rb; Cusabio Technology, LLC.

Techniques: Migration, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Over Expression, CCK-8 Assay