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

Journal: Clinical and Translational Radiation Oncology

Article Title: MonoHER selectively enhances the radiotherapy response in p53 wild-type breast cancer via stabilization of p53

doi: 10.1016/j.ctro.2026.101147

Figure Lengend Snippet: Different protein expression of p-ATM, p- p 53 and p53 in breast cancer cells treated with combination of monoHER (M) and radiation (RT). Representative Western blots of p-ATM, p-p53 and p53 in MCF7 (A), T47D (E) and MCF10A (H) cells. Quantification of p-ATM (B), p-p53 (C) and p53 (D) protein expression in MCF7 cells. Quantification of p-ATM (E), p-p53 (F) and p53 (G) protein expression in T47D cells. Quantification of p-ATM (I), p-p53 (J) and p53 (K) protein expression in MCF10A cells. Data are presented as mean ± SEM from three independent experiments. *p < 0.05, **p < 0.01.

Article Snippet: Membranes were blocked in 5% non-fat milk in TBST for 1 h at RT and incubated overnight at 4°C with primary antibodies against p-ATM (1:2000; Cell Signaling Technology, 5883), p-p53 (1:1000; Cell Signaling Technology, 9284) and p53 (1:1000; Cell Signaling Technology, 9282). α-Tubulin (1:1000, Sigma Aldrich, T7451) or Vinculin (1:1000, Sigma Aldrich, V9131) were used as loading controls.

Techniques: Expressing, Western Blot

Journal: Clinical and Translational Radiation Oncology

Article Title: MonoHER selectively enhances the radiotherapy response in p53 wild-type breast cancer via stabilization of p53

doi: 10.1016/j.ctro.2026.101147

Figure Lengend Snippet: Monoher interacts with p53. Biomolecular interactions of monoHER with wild-type p53 (A) and mutant p53 (B). Docking scores are indicated at the bottom of the image. Representative Western blots and quantification (E) of the CETSA experiment with cell lysates of MCF7 cells, which were treated with DMEM (C) or monoHER (D). Data are presented as mean ± SEM from three independent experiments.

Article Snippet: Membranes were blocked in 5% non-fat milk in TBST for 1 h at RT and incubated overnight at 4°C with primary antibodies against p-ATM (1:2000; Cell Signaling Technology, 5883), p-p53 (1:1000; Cell Signaling Technology, 9284) and p53 (1:1000; Cell Signaling Technology, 9282). α-Tubulin (1:1000, Sigma Aldrich, T7451) or Vinculin (1:1000, Sigma Aldrich, V9131) were used as loading controls.

Techniques: Mutagenesis, Western Blot

Journal: Bioactive Materials

Article Title: On-demand mild photothermal cascade platform reprogramming mitochondrial immunity for tendon rejuvenation

doi: 10.1016/j.bioactmat.2026.01.004

Figure Lengend Snippet: LT-NPs-NIR protects TSPCs against oxidative stress-induced senescence and preserves tenogenic phenotype. (A–D) Immunofluorescence staining for DNA damage (γ-H2AX), proliferation (Ki67), and senescence markers (P16, P53). (E–G) Assessment of stemness (SOX2) and tenogenic differentiation markers (SCX, COL1). (H) Quantitative analysis of the indicated markers. (I) qRT-PCR analysis of SASP-related inflammatory mediators (IL-1β, CXCL10) and matrix-degrading enzymes (MMP3, MMP13). (J) Schematic illustrating the mechanism of ROS scavenging and SASP inhibition. Significance: ns, not significant; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. Snt: senescent cells; Yng: young cells.

Article Snippet: After blocking for 1 h, membranes were incubated overnight at 4 °C with primary antibodies against STING (13647, CST, USA; A21051, Abclonal, China), p-STING (72971, CST, USA; AF7416, Affinity, China), IRF3 (ab68481, Abcam, UK), p-IRF3 (29047, CST, USA), P65 (A22331, Abclonal, China; 8242, CST, USA), p-P65 (AP0124, Abclonal, China), P53 (10442-1-AP, Proteintech, USA), SOX9 (sc-166505, Santa Cruz, USA), BMP-2 (ab284387, abcam, USA), OCN (sc-390877, Santa Cruz, USA), and iNOS (ab178945, Abcam, USA).

Techniques: Immunofluorescence, Staining, Quantitative RT-PCR, Inhibition

Journal: Clinical and Translational Radiation Oncology

Article Title: MonoHER selectively enhances the radiotherapy response in p53 wild-type breast cancer via stabilization of p53

doi: 10.1016/j.ctro.2026.101147

Figure Lengend Snippet: Different protein expression of p-ATM, p- p 53 and p53 in breast cancer cells treated with combination of monoHER (M) and radiation (RT). Representative Western blots of p-ATM, p-p53 and p53 in MCF7 (A), T47D (E) and MCF10A (H) cells. Quantification of p-ATM (B), p-p53 (C) and p53 (D) protein expression in MCF7 cells. Quantification of p-ATM (E), p-p53 (F) and p53 (G) protein expression in T47D cells. Quantification of p-ATM (I), p-p53 (J) and p53 (K) protein expression in MCF10A cells. Data are presented as mean ± SEM from three independent experiments. *p < 0.05, **p < 0.01.

Article Snippet: Membranes were blocked in 5% non-fat milk in TBST for 1 h at RT and incubated overnight at 4°C with primary antibodies against p-ATM (1:2000; Cell Signaling Technology, 5883), p-p53 (1:1000; Cell Signaling Technology, 9284) and p53 (1:1000; Cell Signaling Technology, 9282). α-Tubulin (1:1000, Sigma Aldrich, T7451) or Vinculin (1:1000, Sigma Aldrich, V9131) were used as loading controls.

Techniques: Expressing, Western Blot

Journal: Clinical and Translational Radiation Oncology

Article Title: MonoHER selectively enhances the radiotherapy response in p53 wild-type breast cancer via stabilization of p53

doi: 10.1016/j.ctro.2026.101147

Figure Lengend Snippet: Monoher interacts with p53. Biomolecular interactions of monoHER with wild-type p53 (A) and mutant p53 (B). Docking scores are indicated at the bottom of the image. Representative Western blots and quantification (E) of the CETSA experiment with cell lysates of MCF7 cells, which were treated with DMEM (C) or monoHER (D). Data are presented as mean ± SEM from three independent experiments.

Article Snippet: Membranes were blocked in 5% non-fat milk in TBST for 1 h at RT and incubated overnight at 4°C with primary antibodies against p-ATM (1:2000; Cell Signaling Technology, 5883), p-p53 (1:1000; Cell Signaling Technology, 9284) and p53 (1:1000; Cell Signaling Technology, 9282). α-Tubulin (1:1000, Sigma Aldrich, T7451) or Vinculin (1:1000, Sigma Aldrich, V9131) were used as loading controls.

Techniques: Mutagenesis, Western Blot