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: For mitophagy activation studies, four treatment groups were established: Control; CCCP (10 μM, MCE, 100941) for 72 h; FAC (200 μM) for 72 h; and FAC + CCCP (co-treatment for 72 h).

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: 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: For mitophagy activation studies, four treatment groups were established: Control; CCCP (10 μM, MCE, 100941) for 72 h; FAC (200 μM) for 72 h; and FAC + CCCP (co-treatment for 72 h).

Techniques: Western Blot, Expressing, Marker

Journal: Scientific Reports

Article Title: Ex vivo effects of low dose delayed release rapamycin on agonist-induced platelet aggregation, activation and procoagulant platelet phenotypes in domestic cats

doi: 10.1038/s41598-026-46991-z

Figure Lengend Snippet: Ex vivo measurement of mitochondrial membrane potential in live platelets. Platelets isolated from 8 cats were loaded with tetramethylrhodamine methyl ester (TMRM) before stimulation with thrombin, thrombin/collagen (COL) and thrombin/convulxin (CVX). The extent of mitochondrial membrane potential loss was determined by measuring the percent change (%) in TMRM-positive platelets and fold change (FC) (log10) in TMRM median fluorescence intensity (MFI). ( A ) First, platelets were identified on flow cytometry using forward (FS) and side scatter (SS) properties and ( B ) gating for TMRM-positive platelets were established in fluorescent-minus-one (FMO) controls in thrombin + COL stimulated platelets. ( C ) Histograms demonstrating the loss of TMRM following stimulation with thrombin + CVX (red) compared to the resting (unstimulated) platelets (blue) at baseline. At 24 and 48 h after rapamycin treatment, a graduate increase in TMRM positive platelets was shown demonstrating mitochondrial protective effect despite stimulation with thrombin and CVX. ( D ) Violin plots demonstrating that rapamycin at 3, 24 and 48 h prevented the loss of mitochondrial membrane potential upon stimulation by all three agonists. ( E ) Loss of mitochondria membrane potential was shown to be protected by rapamycin when platelets were treated with carbonyl cyanide 3-chlorophenylhydrazone (CCCP) when the change in TMRM was measured as MFI FC. CCCP-treated platelets served as negative controls. * p < 0.05, ** p < 0.01. Dotted lines represent third quartile, median and first quartile. Width of the plots demonstrate frequency at which values occur.

Article Snippet: Tetramethylrhodamine methyl ester (TMRM) (320nM, Invitrogen, Eugene, OR) was added to PRP and placed in 21% oxygen at 37 °C under gentle rocking for 15 min. TMRM-loaded platelets were subsequently stimulated with bovine alpha thrombin (0.001U/ml, 5 min at 37 °C, Haematologic Technologies, Inc., Essex Junction, VT) prior to further activation with equine type I collagen (COL) (4 μg/ml, CHRONO-LOG Corp., Havertown, PA), or convulxin (CVX) (100 ng/ml, Cayman chemicals, Ann Arbor, MI) for an additional 5 min. Unstimulated platelets served as positive control while platelets treated with either 20 μM carbonyl cyanide 3-chlorophenylhydrazone (CCCP) (Invitrogen, Eugene, OR) or 10 μM A23187 (Millipore Sigma, Burlington, MA) served as negative controls (Figs. A - C).

Techniques: Ex Vivo, Membrane, Isolation, Fluorescence, Flow Cytometry