p53  (Novus Biologicals)

Journal: Aging and Disease

Article Title: Anillin Recedes in p53-Dependent Senescence of Tumor Cells and Reappears in Cells Escaping from Senescence

doi: 10.14336/AD.2025.0402

Figure Lengend Snippet: Analysis of the selected senescence markers and anillin levels in HCT116 p53WT and MCF-7 cells induced to senesce by 1 day-treatment with doxorubicin and collected 5 days after senescence induction. ( A, B ) Densitometric analysis of protein levels in HCT116 p53WT ( A ) and MCF-7 ( B ), n=9; statistical analysis was performed using paired one-tailed t-Student test. Relative protein expression means fold change (in the expression of proteins relative to the expression of GAPDH) vs appropriate control. Boxes: Q1, median, Q3; error bars: Minimum, Maximum. ( C ) Representative images from Western blotting of HCT116 p53WT and MCF-7 cell lysates. ( D, E ) Analysis of fluorescence intensity of anillin (whole nucleus area) and representative images ( F, G ) of control and doxorubicin-treated HCT116 p53WT ( D, F ) and MCF-7 cells ( E, G ), n=3; statistical analysis was performed using Wilcoxon matched-pairs signed-rank test. Data on graphs represent individual values for analyzed cells, median, error bars: Minimum, Maximum. Red – anillin, blue – DAPI stained DNA. Scale 20 µm. Statistical significance relative to control: ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001

Article Snippet: The human HCT116 p53-proficient (referred to as HCT116 p53WT) colon cancer cell line and the breast cancer cell line MCF-7 were obtained from ATCC (HCT116 CCL-247; MCF-7 HTB-22).

Techniques: One-tailed Test, Expressing, Control, Western Blot, Fluorescence, Staining

Journal: Aging and Disease

Article Title: Anillin Recedes in p53-Dependent Senescence of Tumor Cells and Reappears in Cells Escaping from Senescence

doi: 10.14336/AD.2025.0402

Figure Lengend Snippet: Analysis of selected senescence markers and anillin levels in HCT 116 p53KO cells induced to senescence by doxorubicin treatment for 1 day and collected 5 days after senescence induction. ( A ) Densitometric analysis of protein levels in control and doxorubicin-treated HCT116 p53KO based on Western blotting results, n=8; statistical analysis was performed using paired one-tailed t-Student test. Statistical significance is shown relative to control. ( B ) Representative images from Western blotting. ( C ) Comparison of anillin and p53 levels in p53-proficient (HCT116 p53WT and MCF7) and p53-deficient (HCT116 p53KO) cells treated with doxorubicin and analyzed using Western blotting, densitometric analysis (normalized to the level of anillin or p53 in control cells), n=3; statistical analysis was performed using Kruskal-Wallis test. Statistical significance is shown for differences between indicated cell lines. Relative protein expression means fold change (in the expression of proteins relative to the expression of GAPDH) vs appropriate control. ( D ) Representative images of immunostained control and doxorubicin-treated HCT116 p53KO cells. Red – anillin, green – lamin A/C, blue – DAPI stained DNA. Scale 20 µm. Boxes: Q1, median, Q3; error bars: Minimum, Maximum. Statistical significance: **** p ≤ 0.0001

Article Snippet: The human HCT116 p53-proficient (referred to as HCT116 p53WT) colon cancer cell line and the breast cancer cell line MCF-7 were obtained from ATCC (HCT116 CCL-247; MCF-7 HTB-22).

Techniques: Control, Western Blot, One-tailed Test, Comparison, Expressing, Staining

Journal: Aging and Disease

Article Title: Anillin Recedes in p53-Dependent Senescence of Tumor Cells and Reappears in Cells Escaping from Senescence

doi: 10.14336/AD.2025.0402

Figure Lengend Snippet: Correlation between p53 and anillin levels during senescence and the escape from senescence in breast cancer MCF-7 and colon cancer HCT116 p53WT cells (see ). ( A-B ). Representative Western blots showing the levels of anillin and p53 in HCT116 p53WT cells ( A ) and MCF-7 cells ( B ). ( C-F ) The level of anillin and p53 on subsequent days of cell culture after senescence induction by doxorubicin in HCT116 p53WT ( C and E ) and MCF-7 cells ( D and F ) n = 4; statistical analysis was performed using one-way ANOVA followed by post hoc analysis (Tukey’s honest significant difference test; HSD test). Statistical significance of differences between indicated days of treatment: * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001. Boxes: Q1, median, Q3; error bars: Minimum, Maximum. ( G ) The heat map shows the levels of anillin and p53 during senescence and escape from senescence in MCF-7 and HCT116 p53WT cells. Heatmap: Original data points are standardized into z-scores.

Article Snippet: The human HCT116 p53-proficient (referred to as HCT116 p53WT) colon cancer cell line and the breast cancer cell line MCF-7 were obtained from ATCC (HCT116 CCL-247; MCF-7 HTB-22).

Techniques: Western Blot, Cell Culture

Journal: Aging and Disease

Article Title: Anillin Recedes in p53-Dependent Senescence of Tumor Cells and Reappears in Cells Escaping from Senescence

doi: 10.14336/AD.2025.0402

Figure Lengend Snippet: Inverse correlation between ANLN and p53 during senescence and escape from senescence in cancer cells. Downregulation of anillin is a consequence of the induction of p53 due to cellular senescence. After some time, senescent cells resume divisions, which is associated with a decrease in p53 levels and an increase in anillin. Performed with Biorender.

Article Snippet: The human HCT116 p53-proficient (referred to as HCT116 p53WT) colon cancer cell line and the breast cancer cell line MCF-7 were obtained from ATCC (HCT116 CCL-247; MCF-7 HTB-22).

Techniques:

Journal: Redox Biology

Article Title: FTMT-mediated suppression of mitophagy links iron accumulation to osteoporosis

doi: 10.1016/j.redox.2026.104157

Figure Lengend Snippet: Iron accumulation impairs mitophagy, promotes senescence, and suppresses osteogenic differentiation in BMSCs. (a) Schematic diagram of extraction of BMSCs from human femur. (b) Western blot analysis of osteogenic marker proteins (RUNX2, ALP) in BMSCs from normal controls and postmenopausal osteoporosis patients and osteoporosis patients with iron accumulation. (c) Alizarin Red S (ARS) staining of BMSCs treated with increasing concentrations of FAC (0, 50, 100, 200 μM) for 21 days and alkaline phosphatase (ALP) staining of BMSCs treated with increasing concentrations of FAC (0, 50, 100, 200 μM) for 14 days. Scale bar: 50 μm. (d) Western blot analysis of osteogenic markers (RUNX2, ALP) in FAC-treated BMSCs for 5 days. (e) RT-qPCR analysis of osteogenic genes ( Runx2, Alpl, Bglap, Sp7 ) in FAC-treated BMSCs for 72h. (f) KEGG pathway enrichment analysis of differentially expressed genes from RNA sequencing of control and 200 μM FAC-treated BMSCs for 72h. (g, h) Immunofluorescence staining of senescence markers (γ-H2AX, H3K9me3) in FAC-treated BMSCs for 72h. Scale bar: 20 μm. (i) Senescence-associated β-galactosidase (SA-β-gal) staining of FAC-treated BMSCs for 72h. Scale bar: 50 μm. (j) Flow cytometric quantification of SA-β-gal activity in FAC-treated BMSCs for 72h. (k) Western blot analysis of senescence-related proteins (P53, P21, P16) in FAC-treated BMSCs for 72h. (l) Mitophagy assessment by immunofluorescence co-staining with Mitophagy Dye (red) and MitoTracker (green) in FAC-treated BMSCs for 72h. Scale bar: 20 μm. (m) Western blot analysis of mitophagy/autophagy-related proteins (PINK1, PARKIN, P62, LC3) in FAC-treated BMSCs for 72h. (n) Mitochondrial membrane potential (MMP) detection by MT-1 staining in FAC-treated BMSCs for 72h. Scale bar: 30 μm. Data are presented as mean ± SEM; One-way ANOVA (Dunnett's multiple-comparison test); * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Cells were lysed, and proteins were separated by SDS-PAGE, transferred to PVDF membranes (Millipore, IPVH00010), and probed with primary antibodies against: RUNX-2 (Abcam, ab236639), ALP (Affinity, DF6225), P53 (Affinity, AF0879), P21 (Affinity, DF6423), P16 (Abcam, ab51243), PINK1 (HUABIO, ER1706-27), PARKIN (HUABIO, ET1702-60), P62 (Abcam, ab109012), LC3 (NOVUS, NB100-2220), FTMT (Abmart, PC20086S), Phospho-PINK1[Ser228] (Cell Signaling, 89010T), Phospho-PINK1[Ser402] (Absin, abs148820), and GAPDH (Affinity, AF7021).

Techniques: Extraction, Western Blot, Marker, Staining, Quantitative RT-PCR, RNA Sequencing, Control, Immunofluorescence, Activity Assay, Membrane, Comparison

Journal: Redox Biology

Article Title: FTMT-mediated suppression of mitophagy links iron accumulation to osteoporosis

doi: 10.1016/j.redox.2026.104157

Figure Lengend Snippet: Mitophagy activation rescues iron accumulation-induced mitochondrial dysfunction, cellular senescence, and impaired osteogenic differentiation in BMSCs. BMSCs were isolated from normal mice and treated with 200 μM FAC with or without CCCP co-treatment for the same duration in each assay. The time points for the indicated assays were the same as those in . (a) Western blot analysis of mitophagy/autophagy-related proteins (PINK1, PARKIN, P62, LC3). (b, c) Flow cytometric analysis of (b) intracellular ROS and (c) mitochondrial superoxide levels. (d) Mitochondrial membrane potential assessment by MT-1 immunofluorescence staining. Scale bar: 30 μm. (e) Cellular ATP content measurement. (f – i) Immunofluorescence analysis of senescence markers (f, h) γ-H2AX and (g, i) H3K9me3. Scale bar: 40 μm. (j) Western blot analysis of senescence-related proteins (P53, P21, P16). (k) Alizarin Red S (ARS) and alkaline phosphatase (ALP) staining. Scale bar: 50 μm. (l) Western blot analysis of osteogenic marker proteins (RUNX2, ALP). Data are presented as mean ± SEM; One-way ANOVA (Tukey's multiple-comparison test); * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Cells were lysed, and proteins were separated by SDS-PAGE, transferred to PVDF membranes (Millipore, IPVH00010), and probed with primary antibodies against: RUNX-2 (Abcam, ab236639), ALP (Affinity, DF6225), P53 (Affinity, AF0879), P21 (Affinity, DF6423), P16 (Abcam, ab51243), PINK1 (HUABIO, ER1706-27), PARKIN (HUABIO, ET1702-60), P62 (Abcam, ab109012), LC3 (NOVUS, NB100-2220), FTMT (Abmart, PC20086S), Phospho-PINK1[Ser228] (Cell Signaling, 89010T), Phospho-PINK1[Ser402] (Absin, abs148820), and GAPDH (Affinity, AF7021).

Techniques: Activation Assay, Isolation, Western Blot, Membrane, Immunofluorescence, Staining, Marker, Comparison

Journal: Redox Biology

Article Title: FTMT-mediated suppression of mitophagy links iron accumulation to osteoporosis

doi: 10.1016/j.redox.2026.104157

Figure Lengend Snippet: Mitophagy activation alleviates BMSC senescence and restores bone mass in iron-accumulating mice. (a) Representative micro-CT images of distal femoral trabecular bone. (b) Quantitative micro-CT analysis of trabecular bone parameters: Tb.BMD (trabecular bone mineral density), BV/TV (bone volume fraction), BS/TV (bone surface density), and Tb.N (trabecular number). (c) Detection of the serum OCN and P1NP levels from the mice in each group. (d) Histological analysis of tibial sections via H&E staining, toluidine blue staining, and DAPI immunofluorescence from the mice in each group. Scale bar: 250 μm. (e) Detection of the bone formation rate by calcein double labeling from the mice in each group. Scale bar: 20 μm. (f – i) Immunofluorescence analysis of senescence markers (γ-H2AX and H3K9me3) in BMSCs isolated from different treatment groups. Scale bar: 50 μm. (j) Western blot analysis of senescence-related proteins (P53, P21, P16) in BMSCs. (k) Western blot analysis of mitophagy/autophagy-related proteins (PINK1, PARKIN, P62, LC3) in BMSCs. (l) Mitochondrial membrane potential assessment by MT-1 immunofluorescence staining in BMSCs. Scale bar: 50 μm. (m) Cellular ATP content measurement in BMSCs. (n – o) Flow cytometric analysis of (n) intracellular ROS and (o) mitochondrial superoxide levels in BMSCs. Data are presented as mean ± SEM; One-way ANOVA (Tukey's multiple-comparison test); * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Cells were lysed, and proteins were separated by SDS-PAGE, transferred to PVDF membranes (Millipore, IPVH00010), and probed with primary antibodies against: RUNX-2 (Abcam, ab236639), ALP (Affinity, DF6225), P53 (Affinity, AF0879), P21 (Affinity, DF6423), P16 (Abcam, ab51243), PINK1 (HUABIO, ER1706-27), PARKIN (HUABIO, ET1702-60), P62 (Abcam, ab109012), LC3 (NOVUS, NB100-2220), FTMT (Abmart, PC20086S), Phospho-PINK1[Ser228] (Cell Signaling, 89010T), Phospho-PINK1[Ser402] (Absin, abs148820), and GAPDH (Affinity, AF7021).

Techniques: Activation Assay, Micro-CT, Staining, Immunofluorescence, Labeling, Isolation, Western Blot, Membrane, Comparison

Journal: Redox Biology

Article Title: FTMT-mediated suppression of mitophagy links iron accumulation to osteoporosis

doi: 10.1016/j.redox.2026.104157

Figure Lengend Snippet: PINK1 overexpression rescues iron accumulation-induced mitochondrial dysfunction, senescence, and osteogenic impairment in BMSCs. The time points for the indicated assays were the same as those in . (a) Western blot analysis of mitophagy/autophagy-related proteins (PINK1, PARKIN, P62, LC3) in BMSCs transduced with control or PINK1-overexpressing lentivirus followed by FAC treatment. (b) Mitochondrial membrane potential assessment by MT-1 immunofluorescence staining. Scale bar: 50 μm. (c) Cellular ATP content measurement. (d, e) Flow cytometric analysis of (d) intracellular ROS and (e) mitochondrial superoxide levels. (f) Western blot analysis of senescence-related proteins (P53, P21, P16). (g – j) Immunofluorescence analysis of senescence markers (γ-H2AX and H3K9me3). Scale bar: 50 μm. (k, l) Alizarin Red S (ARS) staining and Alkaline phosphatase (ALP) staining. Scale bar: 50 μm. (m) Western blot analysis of osteogenic marker proteins (RUNX2, ALP). (n) RT-qPCR analysis of osteogenic genes ( Runx2, Alpl, Bglap, Sp7 ). Data are presented as mean ± SEM; One-way ANOVA (Tukey's multiple-comparison test); * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Cells were lysed, and proteins were separated by SDS-PAGE, transferred to PVDF membranes (Millipore, IPVH00010), and probed with primary antibodies against: RUNX-2 (Abcam, ab236639), ALP (Affinity, DF6225), P53 (Affinity, AF0879), P21 (Affinity, DF6423), P16 (Abcam, ab51243), PINK1 (HUABIO, ER1706-27), PARKIN (HUABIO, ET1702-60), P62 (Abcam, ab109012), LC3 (NOVUS, NB100-2220), FTMT (Abmart, PC20086S), Phospho-PINK1[Ser228] (Cell Signaling, 89010T), Phospho-PINK1[Ser402] (Absin, abs148820), and GAPDH (Affinity, AF7021).

Techniques: Over Expression, Western Blot, Transduction, Control, Membrane, Immunofluorescence, Staining, Marker, Quantitative RT-PCR, Comparison

Journal: Redox Biology

Article Title: FTMT-mediated suppression of mitophagy links iron accumulation to osteoporosis

doi: 10.1016/j.redox.2026.104157

Figure Lengend Snippet: Impaired mitophagy in BMSCs from osteoporosis patients with iron accumulation. (a) Western blot analysis of senescence-related proteins (P53, P21, P16) in BMSCs from normal controls, postmenopausal osteoporosis patients and osteoporosis patients with iron accumulation. (b) Western blot analysis of mitochondrial ferritin (FTMT) expression levels in BMSCs. (c) Western blot analysis of mitophagy/autophagy-related proteins PINK1, p-PINK1(Ser228), PARKIN, P62, and LC3 in BMSCs. (d) Western blot analysis of mitophagy/autophagy-related proteins PINK1, PARKIN, P62, and LC3 in BMSCs of PMOP and IOP group with or without CCCP intervention. (e) Western blot analysis of senescence-related proteins (P53, P21, P16) in BMSCs of PMOP and IOP group with or without CCCP intervention. (f) Western blot analysis of osteogenic marker proteins (RUNX2, ALP) in BMSCs of PMOP and IOP group with or without CCCP intervention.

Article Snippet: Cells were lysed, and proteins were separated by SDS-PAGE, transferred to PVDF membranes (Millipore, IPVH00010), and probed with primary antibodies against: RUNX-2 (Abcam, ab236639), ALP (Affinity, DF6225), P53 (Affinity, AF0879), P21 (Affinity, DF6423), P16 (Abcam, ab51243), PINK1 (HUABIO, ER1706-27), PARKIN (HUABIO, ET1702-60), P62 (Abcam, ab109012), LC3 (NOVUS, NB100-2220), FTMT (Abmart, PC20086S), Phospho-PINK1[Ser228] (Cell Signaling, 89010T), Phospho-PINK1[Ser402] (Absin, abs148820), and GAPDH (Affinity, AF7021).

Techniques: Western Blot, Expressing, Marker

Journal: Tumour Virus Research

Article Title: HPV18E6 and CDK5 virus-host interaction is a prospective therapeutic target for HPV-positive cervical cancer

doi: 10.1016/j.tvr.2026.200339

Figure Lengend Snippet: A mutualistic association of CDK5 and 18E6 proteins. (A) (i). The represented immunoblot image of CDK5 upregulates the level of 18E6 protein. HEK 293 cells were transfected with an increasing amount of pcDNA3.1: His-CDK5 (1, 2, and 4 μg) and a fixed amount of pcDNA3.1: HA-18E6 (2 μg). (ii). The represented immunoblot image of 18E6 upregulates the level of CDK5 protein. HEK 293 cells were transfected with an increasing amount of pcDNA3.1: HA-18E6 (1, 2, and 4 μg) and pcDNA3.1: His-CDK5 (2 μg). (B) (i). The represented immunoblot image of CDK5 upregulates the level of 18E6 protein in the HeLa cell lines. HeLa cells were transfected with an increasing amount of pcDNA3.1: His-CDK5 (1, 2, 3, and 4 μg). (ii). The represented immunoblot image of 18E6 upregulates the level of CDK5 protein in the SAS cell lines. SAS cells were transfected with an increasing amount of pcDNA3.1: HA-18E6 (1, 2, 3, and 4 μg). (C) (i). The represented immunoblot image of CDK5, E6AP, and p53 after silencing 18E6/E7 expression. Total protein lysates were extracted from HeLa cell lines transfected with siRNA against control (siCtrl) or 18 E6 and E7 (si18 E6/E7) for 72 h. (ii-v) The relative expression levels of 18E6, CDK5, p53, and E6AP relative to β-actin and analyzed using ImageJ and GraphPad Prism (n = 3). All data were presented as means ± standard error of the mean (SEM). (∗, P < 0.05; ∗∗, P < 0.01). (D) The represented immunoblot of CDK5, pCDK5 and HPV 18E6 proteins in the stable expression of HPV 18 in the HGK12 cell line (HPV-null primary keratinocytes cell line), HC: Treatment with 0.2 μM CP681301; LC: Treatment with 0.1 μM CP681301.

Article Snippet: The transferred membranes were incubated overnight at 4 °C with the following primary antibodies: monoclonal rabbit anti-HA (Cell Signaling Technology), monoclonal mouse anti-p53 (Santa Cruz), monoclonal mouse anti-HPV18E6 (Santa Cruz), monoclonal mouse anti-CDK5 (Santa Cruz), and monoclonal mouse anti-β-actin (Santa Cruz).

Techniques: Western Blot, Transfection, Expressing, Control

Journal: Tumour Virus Research

Article Title: HPV18E6 and CDK5 virus-host interaction is a prospective therapeutic target for HPV-positive cervical cancer

doi: 10.1016/j.tvr.2026.200339

Figure Lengend Snippet: CP681301 treatment destabilized E6, reduced E6AP and rescued p53. (A). HeLa and C33A cell lines were treated with CP681301 at the concentrations of 0.6 μM and 1.5 μM for 24 h. The protein extracts were subjected to Western blotting for the detection of CDK5, 18E6, E6AP, p53, and β-actin proteins. (B). CP681301 disrupted the 18E6-E6AP protein complex. The inhibition of CP681301 was detected using the GST-pull down assay, where CP681301 was incubated with the purified GST-18E6 protein and Flag-E6AP protein for 2h. After extensive washing, the bound E6AP protein was detected via Western blotting using an anti-Flag antibody. The immunoblot (IB) on the upper panel shows the interaction of E6AP with GST-18E6, while the lower panel shows the Ponceau S stained of the blot.

Article Snippet: The transferred membranes were incubated overnight at 4 °C with the following primary antibodies: monoclonal rabbit anti-HA (Cell Signaling Technology), monoclonal mouse anti-p53 (Santa Cruz), monoclonal mouse anti-HPV18E6 (Santa Cruz), monoclonal mouse anti-CDK5 (Santa Cruz), and monoclonal mouse anti-β-actin (Santa Cruz).

Techniques: Western Blot, Inhibition, Pull Down Assay, Incubation, Purification, Staining