Journal: Frontiers in Immunology

Article Title: Transcriptome-based profiling of programmed cell death signatures identifies two osteoarthritis subtypes and the hub gene DNM1L

doi: 10.3389/fimmu.2026.1816005

Figure Lengend Snippet: Identification and validation of hub gene. (A, B) Evaluation of the discriminative performance of core gene DNM1L across different datasets. (C) qRT-PCR results of DNM1L in OA patients versus controls. (D, E) Plot of WB results of DNM1L in OA patients and controls. ** Means P<0.01.

Article Snippet: The membrane was incubated with an anti-DNM1L antibody (1:1000; ABclonal, Wuhan, China; catalog #A2586) at 4 °C, followed by a goat anti-rabbit horseradish peroxidase (HRP)-conjugated secondary antibody (1:3000; Proteintech Group, Chicago, USA) at room temperature for 1 hour.

Techniques: Biomarker Discovery, Quantitative RT-PCR

Journal: American journal of physiology. Endocrinology and metabolism

Article Title: Nr4a3 Deficiency Disrupts MEK1–ERK1/2–Drp1 Signaling, Driving Adiposity and Glucose Intolerance in Male Mice

doi: 10.1152/ajpendo.00322.2025

Figure Lengend Snippet: Male inguinal adipose tissue was harvested for mRNA and protein. QPCR was completed for A) DRP1, with a significant decrease being observed in Nr4a3 −/− mice. Western blot B) measuring Serine 616 phosphorylation of Drp1, total Drp1, and GAPDH was completed. Quantification of C) pDRP1 relative to total Drp1 and D) total Drp1 relative to GAPDH was measured. n=5–9, t -test significance * p-value < 0.05.

Article Snippet: The following antibodies were used: P38 (SelleckChem, N23C2, 1:1,000), phospho-P38 (Cell Signaling, 9211, 1:1,000), DRP1 (ABclonal, 1:1,000), phospho-DRP1 (Cell Signaling, 4867S, 1:1,000), ERK1/2 (Cell Signaling, 9102, 1:1,000), phospho-ERK1/2 (Cell Signaling, 9101, 1:1,000), and HSP90 (Santa Cruz, sc-13119, 1:1,000).

Techniques: Expressing, Activation Assay, Western Blot, Phospho-proteomics

Journal: American journal of physiology. Endocrinology and metabolism

Article Title: Nr4a3 Deficiency Disrupts MEK1–ERK1/2–Drp1 Signaling, Driving Adiposity and Glucose Intolerance in Male Mice

doi: 10.1152/ajpendo.00322.2025

Figure Lengend Snippet: Differentiated 3T3-L1 adipocytes transduced with shControl or shNr4a3 were treated with NSC2288155 for 15 minutes and harvested for protein analysis. Western blot A) measuring HSP90, total ERK1/ERK2, and pERK1 (T202/Y204)/pERK2 (T185/Y187) was completed. Quantification of B) pERK1 (T202/Y204)/pERK2 (T185/Y187) relative to HSP90 and C) total ERK1/ERK2 relative to HSP90 were measured. Western blot D) measuring HSP90, total DRP1, pDRP1 (S616), total P38, and pP38 (T180/Y182) was completed. Quantification of E) pDRP1 (S616) relative to HSP90, F) total DRP1 relative to HSP90, G) pP38 (T180/Y182) relative to HSP90, and H) total P38 relative to HSP90 were measured. n=3–4, t-test significance * p-value < 0.05, ** p-value < 0.01, *** p-value < 0.001, **** p-value < 0.0001.

Article Snippet: The following antibodies were used: P38 (SelleckChem, N23C2, 1:1,000), phospho-P38 (Cell Signaling, 9211, 1:1,000), DRP1 (ABclonal, 1:1,000), phospho-DRP1 (Cell Signaling, 4867S, 1:1,000), ERK1/2 (Cell Signaling, 9102, 1:1,000), phospho-ERK1/2 (Cell Signaling, 9101, 1:1,000), and HSP90 (Santa Cruz, sc-13119, 1:1,000).

Techniques: Knockdown, Transduction, Western Blot

Journal: Science Advances

Article Title: The skeletal muscle slow myosin heavy chain regulates mammalian metabolic homeostasis through the NRF2 pathway

doi: 10.1126/sciadv.aed2478

Figure Lengend Snippet: ( A to Db ) Cross sections through the Gas muscle of Pax +/+ , Myh7 fl/+ (A to Bb) and Pax7 icre/+ , Myh7 Δ/fl (C to Db) mice at 2 and 12 months, respectively, labeled by immunofluorescence for MyHC-slow (green), troponin T1 (red), and DAPI (blue). Scale bars, 100 μm (Cb and Db). ( E to H ) SDH staining on Gas muscle sections of Pax +/+ , Myh7 fl/+ (E and G) and Pax7 icre/+ , Myh7 Δ/fl (Ea and Ga) mice and quantification of the percentage SDH activity (F and H) at 2 and 12 months of age, respectively ( n = 5). Scale bars, 100 μm (Ea and Ga). ( I to L ) Representative Western blots for troponin T1, PGC-1α, and α-tubulin (loading control) using Gas muscle protein lysates of Pax +/+ , Myh7 fl/+ , Pax3 cre/+ , Myh7 Δ/fl (P0), and Pax7 icre/+ , Myh7 Δ/fl mice at P0 and 2 and 6 months (I) and their densitometric quantification (J to L) ( n = 4). ( M to P ) Representative Western blots for mitochondrial complexes and α-tubulin (loading control) using protein lysates from the Gas muscle of Pax +/+ , Myh7 fl/+ , Pax3 cre/+ , Myh7 Δ/fl (P0), and Pax7 icre/+ , Myh7 Δ/fl mice at P0 and 6 months (M and O) and their densitometric quantification (N to P) ( n = 4 to 6). ( Q to T ) Representative Western blots for FIS1, pDRP1, DRP1, OPA1, Mitofusin 1, Mitofusin 2, and α-tubulin (loading control) using protein lysates from the Gas muscle of Pax +/+ , Myh7 fl/+ , Pax3 cre/+ , Myh7 Δ/fl (P0), and Pax7 icre/+ , Myh7 Δ/fl mice at P0 and 2 and 6 months (Q) and densitometric quantification (R to T) ( n = 4). ( U and V ) Quantification of ATP levels in the Gas muscle of Pax +/+ , Myh7 fl/+ and Pax7 icre/+ , Myh7 Δ/fl mice at 2 and 12 months of age ( n = 5). Data are presented as means ± SEM. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001, and **** P ≤ 0.0001.

Article Snippet: The antibodies used for Western blotting were as follows: anti-mouse MyHC-IIa antibody (SC-71, Developmental Studies Hybridoma Bank) at 1:10 dilution; anti-mouse MyHC-IIb antibody (BFF3, Developmental Studies Hybridoma Bank) at 1:10; anti-mouse alpha-tubulin antibody (30021290, Cell Signaling Technology) at 1:10,000; anti-rabbit GAPDH antibody (PAB932Mu01, Cloud-Clone Corp) at 1:10,000; anti-rabbit Akt antibody (4641, Cell Signaling Technology) at 1:1000; anti-rabbit P-Akt S473 (4640, Cell Signaling Technology) at 1:1000; anti-rabbit Histone H3 antibody (Ab1741, Abcam) at 1:10,000; anti-rabbit troponin T1 antibody (NBP238855, Novus Biologicals) at 1:5000; anti-rabbit PGC-1α antibody (A12348, ABclonal; 66369, Proteintech; and 2178S, Cell Signaling Technology) at 1:2000, 1:2000, and 1:1000, respectively; anti-mouse total OXPHOS cocktail (ab110413, Abcam) at 1:1000; anti-rabbit DRP1 antibody (A21968, ABclonal) at 1:2000; anti-rabbit FIS1 antibody (A19666, ABclonal) at 1:2000; anti-rabbit mitofusin1 antibody (A9880, ABclonal) at 1:2000; anti-rabbit mitofusin2 antibody (A12771, ABclonal) at 1:2000; anti-rabbit GLUT4 (MA5-17176, Thermo Fisher Scientific) at 1:1000; anti-rabbit NRF2 antibody (A0674, ABclonal and 98020000, Proteintech) at 1:2000; anti-rabbit P-NRF2 antibody (AP1133, ABclonal) at 1:2000; anti-rabbit SOD1 antibody (A0274, ABclonal) at 1:2000; anti-rabbit Prdx1 antibody (A13524, ABclonal) at 1:2000; anti-rabbit Gpx1 antibody (A1110, ABclonal) at 1:1000; anti-rabbit catalase antibody (A11220, ABclonal) at 1:2000; anti-rabbit KEAP1 antibody (A1820, ABclonal) at 1:1000; anti-rabbit Tom20 antibody (HPA011582, Merck) at 1:2500, anti-rabbit VDAC1 antibody ( Ab154856 , Abcam) at 1:2500; anti-rabbit pDRP1 (S616) antibody (AP0849, ABclonal) at 1:2500; and anti-rabbit OPA1 antibody (A9833, ABclonal) at 1:2500.

Techniques: Labeling, Immunofluorescence, Staining, Activity Assay, Western Blot, Control

Journal: Materials Today Bio

Article Title: A dual-responsive CO-releasing nanogel ameliorates retinal ischemia–reperfusion injury by restoring mitochondrial homeostasis and attenuating cGAS-STING pathway activation

doi: 10.1016/j.mtbio.2026.102974

Figure Lengend Snippet: COPN restores mitochondrial quality control and interrupts the ROS‒cGAS‒inflammation axis in OGD/R-treated R28 cells. a , TEM images showing changes in the mitochondrial ultrastructure of R28 cells subjected to various treatments (PBS, PDNs, COPN-L, and COPN-H) after OGD/R. Scale bar: 500 nm b , Quantitative analyses of mitochondrial length and number in R28 cells as indicated in a . c , Representative fluorescence images showing JC-1 staining of the mitochondrial Δψm: aggregates (red) indicate healthy mitochondria, whereas monomers (green) represent depolarized mitochondria. Nuclei were stained with DAPI (blue). Scale bar: 50 μm d , Intracellular ATP content assay revealing improved energy production after COPN treatment under OGD/R stress conditions. e , mRNA expression analysis of mitochondrial dynamic regulatory genes (Opa1, Mfn2, Drp1, and Fis1) and mitochondrial DNA transcription levels (mt-ND1 and mt-COX1). f , Quantitative analysis of the JC-1 fluorescence ratio (aggregates/monomers) and mitochondrial ROS levels (MitoSOX staining). g–h , Western blot analysis of proteins involved in mitochondrial autophagy (Pink1, Parkin, and P62) (g) and mitochondrial fusion/fission (Opa1, Mfn2, Mfn1, Drp1, and Fis1) (h) . i , Confocal fluorescence microscopy images of mitochondria (green, MitoTracker) and lysosomes (red, LysoTracker) in R28 cells after different treatments. Nuclei were stained with Hoechst (blue). Scale bar: 50 μm ∗ ∗∗∗P < 0.0001. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The following primary antibodies were used: Drp1 (CST, #8570, 1:1000, ∼80 kDa), Fis1 (Proteintech, 10956-1-AP, 1:1000, ∼17 kDa), Mfn1 (Abcam, ab104274, 1:1000, ∼84 kDa), Mfn2 (CST, #9482, 1:1000, ∼86 kDa), Opa1 (CST, #80471, 1:1000, ∼100–120 kDa), Pink1 (CST, #6946, 1:1000, ∼63 kDa), cGAS (CST, #15102, 1:1000, ∼60 kDa), STING (CST, #13647, 1:1000, ∼42–45 kDa), TBK1 (CST, #3013, 1:1000, ∼84 kDa), p-TBK1 (CST, #5483, 1:1000, ∼84 kDa), COX IV (Abcam, ab14744, 1:2000, ∼17 kDa), β-actin (Proteintech, 66009-1-Ig, 1:5000, ∼43 kDa), and GAPDH (Proteintech, 60004-1-Ig, 1:5000, After washing, the membranes were incubated with HRP-conjugated secondary antibodies (goat anti-rabbit or goat anti-mouse IgG, CST, 1:5000) for 1 h at room temperature.

Techniques: Control, Fluorescence, Staining, Expressing, Western Blot, Microscopy

Journal: International Journal of Molecular Medicine

Article Title: ZEB1 maintains mitochondrial fission and macrophage efferocytosis by restraining MFN2, thereby limiting inflammation and improving tendon-bone healing

doi: 10.3892/ijmm.2026.5805

Figure Lengend Snippet: ZEB1 affects mitochondrial fission by regulating MFN2. (A) Confocal microscopy results of mitochondrial morphology in macrophages. Mitochondria were labeled with Mito-Tracker Red fluorescent probe (scale bar, 10 µ m; lower panels, ×200 magnification). Western blotting analysis of (B) MFN2 and (C) DRP1 protein expression in macrophages after ZEB1 knockdown. Quantitative density analysis data are presented as mean ± standard deviation (n=3 independent experiments). (D) Immunofluorescence colocalization analysis of MFN2 localization in mitochondria. Nuclei were stained with DAPI (blue), MFN2 protein was labeled with Alexa Fluor 488 (green) and mitochondrial membrane protein TOMM20 was labeled with Alexa Fluor 594 (red) (scale bar, 10 µ m). n=3, * P<0.05 vs. NC. DRP1, dynamin-related protein 1; MFN2, Mitofusin-2; ZEB1, zinc finger E-box binding homeobox 1; NC, negative control; sh, short hairpin.

Article Snippet: The primary antibodies used were as follows: Anti-β-Actin antibody (1:5,000; cat. no. 81115-1-RR; Proteintech Group, Inc.), anti-ZEB1 antibody (1:1,000; cat. no. ab203829; Abcam), anti-MFN2 antibody (1:5,000; cat. no. ab124773; Abcam), and anti-DRP1 antibody (1:5,000; cat. no. 5391; Cell Signaling Technology, Inc.).

Techniques: Confocal Microscopy, Labeling, Western Blot, Expressing, Knockdown, Standard Deviation, Immunofluorescence, Staining, Membrane, Binding Assay, Negative Control