Journal: Materials Today Bio

Article Title: A biomimetic multimodal nanoplatform combining neutrophil-coated two-dimensional metalloporphyrinic framework nanosheet and exendin-4 to treat obesity-related osteoporosis

doi: 10.1016/j.mtbio.2025.102009

Figure Lengend Snippet: The enhancement of Ex-4 on osteoblasts within a Pa environment is associated with mitophagy. ( A ) and ( F ) depict intracellular reactive oxygen species (ROS) levels using Dichlorodihydrofluorescein diacetate (DCFH-DA) (green) staining, with quantitative analysis provided. Scale bar: 100 μm. ( B ) It illustrates the alterations in mitochondrial morphology across different environments, as marked by MitoTracker. Scale bar: 20 μm. ( C ) and ( G ) Current alterations in mitochondrial membrane potential (ΔΨm) are identified using the JC-1 fluorescent probe. Scale bar: 30 μm. ( D ) and ( H ) It assesses the colocalization levels of mitochondria and lysosomes in various environments, with COX IV (red), LAMP1 (green), and DAPI (blue) marking mitochondria, lysosomes, and nuclei, respectively. Scale bar: 20 μm. ( E ) It employs transmission electron microscopy (TEM) to observe the formation of mitochondria and autophagosomes in osteoblasts under different conditions. Scale bar: 2 μm. ( I-J ) It utilizes WB analysis to examine autophagy-related proteins ATG4B and LC3B. Results are expressed as Mean ± SD, with significance denoted by ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001, n = 3.

Article Snippet: Items and chemicals utilized in this experiment: MC3T3-E1 subclone 24 (EallBio, China), Alpha modification of Eagle's medium (α-MEM, EallBio, China), Fetal bovine serum (FBS, CELL-BOX, China), Trypsin (NCM, China), A mixture of protease and phosphatase inhibitors (NCM, China), Phosphate-buffered saline (PBS, EallBio, China), Bovine Serum Albumin (BSA, Solarbio, China), Cell Counting Kit-8 (CCK-8, NCM, China), Bicin-choninic Acid (BCA, NCM, China), Enhanced chemiluminescent reagent (NCM, China), RIPA assay buffer (NCM, China), Blotting reagent (NCM, China), Color page gel rapid preparation kit (Epizyme, USA), Multicolor prestained protein ladder (Epizyme, USA), Mitotracker Red CMXRos (Beyotime, China), Mitochondrial membrane potential (ΔΨm) assay kit with JC-1 (Beyotime, China), ROS kit (Beyotime, China), 4′,6-Diamidino-2-phenylindole (DAPI, Beyotime, China), Hoechst (Beyotime, China), 4 % Paraformaldehyde (PFA, Solarbio, China), sodium citrate buffer (Solarbio, China), formalin-EDTA decalcifying solution (Solarbio, China), Hematoxylin-eosin stain kit (H&E, Solarbio, China), Modified Masson's trichrome stain kit (Masson, Solarbio, China), Triton X-100 (Servicebio, China), Enhanced endogenous peroxidase blocking buffer (Beyotime, China), Diaminobenzidine (DAB, Zsbio, China), Tris buffered saline with Tween-20 (TBST, Solarbio, China), TRIzol (Sigma, USA), ALN (Sigma, USA), Ex-4 (Selleck, USA), Dexamethasone (MCE, USA), β-Glycerophosphate disodium salt pentahydrate (MCE, USA) and Vitamin C (MCE, USA), Palmitic acid (Pa, Kunchuang Technology, China), Bafilomycin A1 (BafA1, MCE, USA).

Techniques: Staining, Membrane, Transmission Assay, Electron Microscopy

Journal: Materials Today Bio

Article Title: A biomimetic multimodal nanoplatform combining neutrophil-coated two-dimensional metalloporphyrinic framework nanosheet and exendin-4 to treat obesity-related osteoporosis

doi: 10.1016/j.mtbio.2025.102009

Figure Lengend Snippet: DPA@NM@CTZE reduces excessive mitophagy, enhancing osteoblast function under high lipid conditions. ( A ) and ( F ) show changes in intracellular ROS levels post-treatment with various materials. Scale bar: 100 μm. ( B ) Displays mitochondrial morphological changes in treated osteoblasts. Scale bar: 20 μm. ( C ) and ( G ) JC-1 detection of the improvement in ΔΨm by DPA@NM@CTZE. Scale bar: 30 μm. ( D ) and ( H ) The effects of different materials on osteoblasts in a Pa environment were analyzed by assessing the degree of co-localization of mitochondria and lysosomes. Scale bar: 20 μm. ( E ) The effects of different material components on mitochondria and autophagosomes were observed at the microscopic level using TEM. Scale bar: 2 μm. ( I-J ) WB was used to detect the changes in PINK1/Parkin autophagy pathway, the autophagy-related proteins and osteoblast proteins. Data are presented as Mean ± SD. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, n = 3.

Article Snippet: Items and chemicals utilized in this experiment: MC3T3-E1 subclone 24 (EallBio, China), Alpha modification of Eagle's medium (α-MEM, EallBio, China), Fetal bovine serum (FBS, CELL-BOX, China), Trypsin (NCM, China), A mixture of protease and phosphatase inhibitors (NCM, China), Phosphate-buffered saline (PBS, EallBio, China), Bovine Serum Albumin (BSA, Solarbio, China), Cell Counting Kit-8 (CCK-8, NCM, China), Bicin-choninic Acid (BCA, NCM, China), Enhanced chemiluminescent reagent (NCM, China), RIPA assay buffer (NCM, China), Blotting reagent (NCM, China), Color page gel rapid preparation kit (Epizyme, USA), Multicolor prestained protein ladder (Epizyme, USA), Mitotracker Red CMXRos (Beyotime, China), Mitochondrial membrane potential (ΔΨm) assay kit with JC-1 (Beyotime, China), ROS kit (Beyotime, China), 4′,6-Diamidino-2-phenylindole (DAPI, Beyotime, China), Hoechst (Beyotime, China), 4 % Paraformaldehyde (PFA, Solarbio, China), sodium citrate buffer (Solarbio, China), formalin-EDTA decalcifying solution (Solarbio, China), Hematoxylin-eosin stain kit (H&E, Solarbio, China), Modified Masson's trichrome stain kit (Masson, Solarbio, China), Triton X-100 (Servicebio, China), Enhanced endogenous peroxidase blocking buffer (Beyotime, China), Diaminobenzidine (DAB, Zsbio, China), Tris buffered saline with Tween-20 (TBST, Solarbio, China), TRIzol (Sigma, USA), ALN (Sigma, USA), Ex-4 (Selleck, USA), Dexamethasone (MCE, USA), β-Glycerophosphate disodium salt pentahydrate (MCE, USA) and Vitamin C (MCE, USA), Palmitic acid (Pa, Kunchuang Technology, China), Bafilomycin A1 (BafA1, MCE, USA).

Techniques:

Journal: Materials Today Bio

Article Title: A biomimetic multimodal nanoplatform combining neutrophil-coated two-dimensional metalloporphyrinic framework nanosheet and exendin-4 to treat obesity-related osteoporosis

doi: 10.1016/j.mtbio.2025.102009

Figure Lengend Snippet: DPA@NM@CTZE enhances osteoblast function by inhibiting the PINK1/Parkin signaling pathway. ( A ) and ( F ) Show the impact of LV-sh Pink1 transfection on ROS levels in osteoblast. scale bar: 100 μm. ( B ) Illustrates the effects of LV-sh Pink1 and DPA@NM@CTZE on mitochondrial morphology. scale bar: 20 μm. ( C ) and ( G ) Indicate that inhibiting the PINK1/Parkin pathway increases ΔΨm scale bar: 30 μm. ( D ) and ( H ) To prevent self-fluorescence post-LV-sh Pink1 transfection, mitochondria, lysosomes, and the nucleus were labeled with COX IV (red), LAMP1 (yellow), and DAPI (blue), respectively, to observe mitochondria-lysosome colocalization. Scale bar: 20 μm. ( E ) To examine mitochondrial and autophagosome changes post-transfection. Scale indicator: 2 μm. ( I-J ) Impact of LV-sh Pink1 transfection on mitophagy and osteogenic proteins. Data as Mean ± SD. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, n = 3.

Article Snippet: Items and chemicals utilized in this experiment: MC3T3-E1 subclone 24 (EallBio, China), Alpha modification of Eagle's medium (α-MEM, EallBio, China), Fetal bovine serum (FBS, CELL-BOX, China), Trypsin (NCM, China), A mixture of protease and phosphatase inhibitors (NCM, China), Phosphate-buffered saline (PBS, EallBio, China), Bovine Serum Albumin (BSA, Solarbio, China), Cell Counting Kit-8 (CCK-8, NCM, China), Bicin-choninic Acid (BCA, NCM, China), Enhanced chemiluminescent reagent (NCM, China), RIPA assay buffer (NCM, China), Blotting reagent (NCM, China), Color page gel rapid preparation kit (Epizyme, USA), Multicolor prestained protein ladder (Epizyme, USA), Mitotracker Red CMXRos (Beyotime, China), Mitochondrial membrane potential (ΔΨm) assay kit with JC-1 (Beyotime, China), ROS kit (Beyotime, China), 4′,6-Diamidino-2-phenylindole (DAPI, Beyotime, China), Hoechst (Beyotime, China), 4 % Paraformaldehyde (PFA, Solarbio, China), sodium citrate buffer (Solarbio, China), formalin-EDTA decalcifying solution (Solarbio, China), Hematoxylin-eosin stain kit (H&E, Solarbio, China), Modified Masson's trichrome stain kit (Masson, Solarbio, China), Triton X-100 (Servicebio, China), Enhanced endogenous peroxidase blocking buffer (Beyotime, China), Diaminobenzidine (DAB, Zsbio, China), Tris buffered saline with Tween-20 (TBST, Solarbio, China), TRIzol (Sigma, USA), ALN (Sigma, USA), Ex-4 (Selleck, USA), Dexamethasone (MCE, USA), β-Glycerophosphate disodium salt pentahydrate (MCE, USA) and Vitamin C (MCE, USA), Palmitic acid (Pa, Kunchuang Technology, China), Bafilomycin A1 (BafA1, MCE, USA).

Techniques: Transfection, Fluorescence, Labeling

Journal: Materials Today Bio

Article Title: A biomimetic multimodal nanoplatform combining neutrophil-coated two-dimensional metalloporphyrinic framework nanosheet and exendin-4 to treat obesity-related osteoporosis

doi: 10.1016/j.mtbio.2025.102009

Figure Lengend Snippet: TET2 plays a crucial role in DPA@NM@CTZE's inhibition of autophagosome formation. ( A ) Assess the impact of reduced Tet2 and Bcl2 expression with DPA@NM@CTZE on intracellular ROS levels. Scale bar: 100 μm. ( B ) Examine how LV-sh Tet2 and si- Bcl2 transfection affects mitochondrial morphology Scale bar: 20 μm. ( C ) Evaluate the influence of transfected cells, with or without LV-sh Tet2 and si- Bcl2 , on osteoblasts' cell ΔΨm. Scale bar: 30 μm. ( D ) Colocalization images show mitochondria (red) and lysosomes (yellow) in LV-sh Tet2 and si- Bcl2 transfected cells. Scale: 20 μm. ( E ) TEM analysis reveals the impact of reduced Tet2 and Bcl2 on mitochondria and autophagosomes in osteoblasts. Scale: 2 μm. (F–G) WB assess the effects of decreased Tet2 and Bcl2 on mitophagy and osteoblast protein expression. Data: Mean ± SD. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, n = 3.

Article Snippet: Items and chemicals utilized in this experiment: MC3T3-E1 subclone 24 (EallBio, China), Alpha modification of Eagle's medium (α-MEM, EallBio, China), Fetal bovine serum (FBS, CELL-BOX, China), Trypsin (NCM, China), A mixture of protease and phosphatase inhibitors (NCM, China), Phosphate-buffered saline (PBS, EallBio, China), Bovine Serum Albumin (BSA, Solarbio, China), Cell Counting Kit-8 (CCK-8, NCM, China), Bicin-choninic Acid (BCA, NCM, China), Enhanced chemiluminescent reagent (NCM, China), RIPA assay buffer (NCM, China), Blotting reagent (NCM, China), Color page gel rapid preparation kit (Epizyme, USA), Multicolor prestained protein ladder (Epizyme, USA), Mitotracker Red CMXRos (Beyotime, China), Mitochondrial membrane potential (ΔΨm) assay kit with JC-1 (Beyotime, China), ROS kit (Beyotime, China), 4′,6-Diamidino-2-phenylindole (DAPI, Beyotime, China), Hoechst (Beyotime, China), 4 % Paraformaldehyde (PFA, Solarbio, China), sodium citrate buffer (Solarbio, China), formalin-EDTA decalcifying solution (Solarbio, China), Hematoxylin-eosin stain kit (H&E, Solarbio, China), Modified Masson's trichrome stain kit (Masson, Solarbio, China), Triton X-100 (Servicebio, China), Enhanced endogenous peroxidase blocking buffer (Beyotime, China), Diaminobenzidine (DAB, Zsbio, China), Tris buffered saline with Tween-20 (TBST, Solarbio, China), TRIzol (Sigma, USA), ALN (Sigma, USA), Ex-4 (Selleck, USA), Dexamethasone (MCE, USA), β-Glycerophosphate disodium salt pentahydrate (MCE, USA) and Vitamin C (MCE, USA), Palmitic acid (Pa, Kunchuang Technology, China), Bafilomycin A1 (BafA1, MCE, USA).

Techniques: Inhibition, Expressing, Transfection

Journal: Theranostics

Article Title: Preclinical activity of DCZ3301, a novel aryl-guanidino compound in the therapy of multiple myeloma

doi: 10.7150/thno.18345

Figure Lengend Snippet: DCZ3301 induced apoptosis and decreased ΔΨm in MM cells. (A) MM.1S cells were treated with DCZ3301 (0-8 μM) for 48 and 72 h. Cell apoptosis was analyzed by flow cytometry using Annexin V/PI staining. Columns (right panel) represent the average percent of Annexin V positive cells from three independent experiments, which are shown as the mean±SD. (B) MM.1S cells were treated with DCZ3301 (0-8 μM) for 48 h. Whole cell lysates were subjected to western blotting using caspase 8, caspase 9, caspase 3, PARP and β-actin. (C) MM.1S cells were incubated with or without pan-caspase inhibitor Z-VAD-FMK for 1 h and then treated with DCZ3301 (4 μM) for 48 h followed by assessment of cell apoptosis using Annexin V/PI staining. Columns (right panel) represent the average percent of Annexin V positive cells from three independent experiments, which are shown as the mean±SD. (D) MM.1S cells were treated with DCZ3301 (0-8 μM) for 48 h. Mitochondrial membrane potential (ΔΨm) was analyzed by flow cytometry using JC-1 dye. Only JC-1 green positive (lower right quadrant) cells were analyzed for the loss of ΔΨm. Columns (lower panel) represent the average percent of only JC-1 green positive cells from three independent experiments, which are shown as the mean±SD. # represent p >0.05; * represent p <0.05; * * represent p <0.01; * * *represent p <0.001.

Article Snippet: Mitochondrial membrane potentials (ΔΨm) were measured using JC-1 dye (Beyotime Institute of Biotechnology), as previously described .

Techniques: Flow Cytometry, Staining, Western Blot, Incubation, Membrane

Journal: Materials Today Bio

Article Title: A biomimetic multimodal nanoplatform combining neutrophil-coated two-dimensional metalloporphyrinic framework nanosheet and exendin-4 to treat obesity-related osteoporosis

doi: 10.1016/j.mtbio.2025.102009

Figure Lengend Snippet: The enhancement of Ex-4 on osteoblasts within a Pa environment is associated with mitophagy. ( A ) and ( F ) depict intracellular reactive oxygen species (ROS) levels using Dichlorodihydrofluorescein diacetate (DCFH-DA) (green) staining, with quantitative analysis provided. Scale bar: 100 μm. ( B ) It illustrates the alterations in mitochondrial morphology across different environments, as marked by MitoTracker. Scale bar: 20 μm. ( C ) and ( G ) Current alterations in mitochondrial membrane potential (ΔΨm) are identified using the JC-1 fluorescent probe. Scale bar: 30 μm. ( D ) and ( H ) It assesses the colocalization levels of mitochondria and lysosomes in various environments, with COX IV (red), LAMP1 (green), and DAPI (blue) marking mitochondria, lysosomes, and nuclei, respectively. Scale bar: 20 μm. ( E ) It employs transmission electron microscopy (TEM) to observe the formation of mitochondria and autophagosomes in osteoblasts under different conditions. Scale bar: 2 μm. ( I-J ) It utilizes WB analysis to examine autophagy-related proteins ATG4B and LC3B. Results are expressed as Mean ± SD, with significance denoted by ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001, n = 3.

Article Snippet: Items and chemicals utilized in this experiment: MC3T3-E1 subclone 24 (EallBio, China), Alpha modification of Eagle's medium (α-MEM, EallBio, China), Fetal bovine serum (FBS, CELL-BOX, China), Trypsin (NCM, China), A mixture of protease and phosphatase inhibitors (NCM, China), Phosphate-buffered saline (PBS, EallBio, China), Bovine Serum Albumin (BSA, Solarbio, China), Cell Counting Kit-8 (CCK-8, NCM, China), Bicin-choninic Acid (BCA, NCM, China), Enhanced chemiluminescent reagent (NCM, China), RIPA assay buffer (NCM, China), Blotting reagent (NCM, China), Color page gel rapid preparation kit (Epizyme, USA), Multicolor prestained protein ladder (Epizyme, USA), Mitotracker Red CMXRos (Beyotime, China), Mitochondrial membrane potential (ΔΨm) assay kit with JC-1 (Beyotime, China), ROS kit (Beyotime, China), 4′,6-Diamidino-2-phenylindole (DAPI, Beyotime, China), Hoechst (Beyotime, China), 4 % Paraformaldehyde (PFA, Solarbio, China), sodium citrate buffer (Solarbio, China), formalin-EDTA decalcifying solution (Solarbio, China), Hematoxylin-eosin stain kit (H&E, Solarbio, China), Modified Masson's trichrome stain kit (Masson, Solarbio, China), Triton X-100 (Servicebio, China), Enhanced endogenous peroxidase blocking buffer (Beyotime, China), Diaminobenzidine (DAB, Zsbio, China), Tris buffered saline with Tween-20 (TBST, Solarbio, China), TRIzol (Sigma, USA), ALN (Sigma, USA), Ex-4 (Selleck, USA), Dexamethasone (MCE, USA), β-Glycerophosphate disodium salt pentahydrate (MCE, USA) and Vitamin C (MCE, USA), Palmitic acid (Pa, Kunchuang Technology, China), Bafilomycin A1 (BafA1, MCE, USA).

Techniques: Staining, Membrane, Transmission Assay, Electron Microscopy

Journal: Materials Today Bio

Article Title: A biomimetic multimodal nanoplatform combining neutrophil-coated two-dimensional metalloporphyrinic framework nanosheet and exendin-4 to treat obesity-related osteoporosis

doi: 10.1016/j.mtbio.2025.102009

Figure Lengend Snippet: DPA@NM@CTZE reduces excessive mitophagy, enhancing osteoblast function under high lipid conditions. ( A ) and ( F ) show changes in intracellular ROS levels post-treatment with various materials. Scale bar: 100 μm. ( B ) Displays mitochondrial morphological changes in treated osteoblasts. Scale bar: 20 μm. ( C ) and ( G ) JC-1 detection of the improvement in ΔΨm by DPA@NM@CTZE. Scale bar: 30 μm. ( D ) and ( H ) The effects of different materials on osteoblasts in a Pa environment were analyzed by assessing the degree of co-localization of mitochondria and lysosomes. Scale bar: 20 μm. ( E ) The effects of different material components on mitochondria and autophagosomes were observed at the microscopic level using TEM. Scale bar: 2 μm. ( I-J ) WB was used to detect the changes in PINK1/Parkin autophagy pathway, the autophagy-related proteins and osteoblast proteins. Data are presented as Mean ± SD. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, n = 3.

Article Snippet: Items and chemicals utilized in this experiment: MC3T3-E1 subclone 24 (EallBio, China), Alpha modification of Eagle's medium (α-MEM, EallBio, China), Fetal bovine serum (FBS, CELL-BOX, China), Trypsin (NCM, China), A mixture of protease and phosphatase inhibitors (NCM, China), Phosphate-buffered saline (PBS, EallBio, China), Bovine Serum Albumin (BSA, Solarbio, China), Cell Counting Kit-8 (CCK-8, NCM, China), Bicin-choninic Acid (BCA, NCM, China), Enhanced chemiluminescent reagent (NCM, China), RIPA assay buffer (NCM, China), Blotting reagent (NCM, China), Color page gel rapid preparation kit (Epizyme, USA), Multicolor prestained protein ladder (Epizyme, USA), Mitotracker Red CMXRos (Beyotime, China), Mitochondrial membrane potential (ΔΨm) assay kit with JC-1 (Beyotime, China), ROS kit (Beyotime, China), 4′,6-Diamidino-2-phenylindole (DAPI, Beyotime, China), Hoechst (Beyotime, China), 4 % Paraformaldehyde (PFA, Solarbio, China), sodium citrate buffer (Solarbio, China), formalin-EDTA decalcifying solution (Solarbio, China), Hematoxylin-eosin stain kit (H&E, Solarbio, China), Modified Masson's trichrome stain kit (Masson, Solarbio, China), Triton X-100 (Servicebio, China), Enhanced endogenous peroxidase blocking buffer (Beyotime, China), Diaminobenzidine (DAB, Zsbio, China), Tris buffered saline with Tween-20 (TBST, Solarbio, China), TRIzol (Sigma, USA), ALN (Sigma, USA), Ex-4 (Selleck, USA), Dexamethasone (MCE, USA), β-Glycerophosphate disodium salt pentahydrate (MCE, USA) and Vitamin C (MCE, USA), Palmitic acid (Pa, Kunchuang Technology, China), Bafilomycin A1 (BafA1, MCE, USA).

Techniques:

Journal: Materials Today Bio

Article Title: A biomimetic multimodal nanoplatform combining neutrophil-coated two-dimensional metalloporphyrinic framework nanosheet and exendin-4 to treat obesity-related osteoporosis

doi: 10.1016/j.mtbio.2025.102009

Figure Lengend Snippet: DPA@NM@CTZE enhances osteoblast function by inhibiting the PINK1/Parkin signaling pathway. ( A ) and ( F ) Show the impact of LV-sh Pink1 transfection on ROS levels in osteoblast. scale bar: 100 μm. ( B ) Illustrates the effects of LV-sh Pink1 and DPA@NM@CTZE on mitochondrial morphology. scale bar: 20 μm. ( C ) and ( G ) Indicate that inhibiting the PINK1/Parkin pathway increases ΔΨm scale bar: 30 μm. ( D ) and ( H ) To prevent self-fluorescence post-LV-sh Pink1 transfection, mitochondria, lysosomes, and the nucleus were labeled with COX IV (red), LAMP1 (yellow), and DAPI (blue), respectively, to observe mitochondria-lysosome colocalization. Scale bar: 20 μm. ( E ) To examine mitochondrial and autophagosome changes post-transfection. Scale indicator: 2 μm. ( I-J ) Impact of LV-sh Pink1 transfection on mitophagy and osteogenic proteins. Data as Mean ± SD. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, n = 3.

Article Snippet: Items and chemicals utilized in this experiment: MC3T3-E1 subclone 24 (EallBio, China), Alpha modification of Eagle's medium (α-MEM, EallBio, China), Fetal bovine serum (FBS, CELL-BOX, China), Trypsin (NCM, China), A mixture of protease and phosphatase inhibitors (NCM, China), Phosphate-buffered saline (PBS, EallBio, China), Bovine Serum Albumin (BSA, Solarbio, China), Cell Counting Kit-8 (CCK-8, NCM, China), Bicin-choninic Acid (BCA, NCM, China), Enhanced chemiluminescent reagent (NCM, China), RIPA assay buffer (NCM, China), Blotting reagent (NCM, China), Color page gel rapid preparation kit (Epizyme, USA), Multicolor prestained protein ladder (Epizyme, USA), Mitotracker Red CMXRos (Beyotime, China), Mitochondrial membrane potential (ΔΨm) assay kit with JC-1 (Beyotime, China), ROS kit (Beyotime, China), 4′,6-Diamidino-2-phenylindole (DAPI, Beyotime, China), Hoechst (Beyotime, China), 4 % Paraformaldehyde (PFA, Solarbio, China), sodium citrate buffer (Solarbio, China), formalin-EDTA decalcifying solution (Solarbio, China), Hematoxylin-eosin stain kit (H&E, Solarbio, China), Modified Masson's trichrome stain kit (Masson, Solarbio, China), Triton X-100 (Servicebio, China), Enhanced endogenous peroxidase blocking buffer (Beyotime, China), Diaminobenzidine (DAB, Zsbio, China), Tris buffered saline with Tween-20 (TBST, Solarbio, China), TRIzol (Sigma, USA), ALN (Sigma, USA), Ex-4 (Selleck, USA), Dexamethasone (MCE, USA), β-Glycerophosphate disodium salt pentahydrate (MCE, USA) and Vitamin C (MCE, USA), Palmitic acid (Pa, Kunchuang Technology, China), Bafilomycin A1 (BafA1, MCE, USA).

Techniques: Transfection, Fluorescence, Labeling

Journal: Materials Today Bio

Article Title: A biomimetic multimodal nanoplatform combining neutrophil-coated two-dimensional metalloporphyrinic framework nanosheet and exendin-4 to treat obesity-related osteoporosis

doi: 10.1016/j.mtbio.2025.102009

Figure Lengend Snippet: TET2 plays a crucial role in DPA@NM@CTZE's inhibition of autophagosome formation. ( A ) Assess the impact of reduced Tet2 and Bcl2 expression with DPA@NM@CTZE on intracellular ROS levels. Scale bar: 100 μm. ( B ) Examine how LV-sh Tet2 and si- Bcl2 transfection affects mitochondrial morphology Scale bar: 20 μm. ( C ) Evaluate the influence of transfected cells, with or without LV-sh Tet2 and si- Bcl2 , on osteoblasts' cell ΔΨm. Scale bar: 30 μm. ( D ) Colocalization images show mitochondria (red) and lysosomes (yellow) in LV-sh Tet2 and si- Bcl2 transfected cells. Scale: 20 μm. ( E ) TEM analysis reveals the impact of reduced Tet2 and Bcl2 on mitochondria and autophagosomes in osteoblasts. Scale: 2 μm. (F–G) WB assess the effects of decreased Tet2 and Bcl2 on mitophagy and osteoblast protein expression. Data: Mean ± SD. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, n = 3.

Article Snippet: Items and chemicals utilized in this experiment: MC3T3-E1 subclone 24 (EallBio, China), Alpha modification of Eagle's medium (α-MEM, EallBio, China), Fetal bovine serum (FBS, CELL-BOX, China), Trypsin (NCM, China), A mixture of protease and phosphatase inhibitors (NCM, China), Phosphate-buffered saline (PBS, EallBio, China), Bovine Serum Albumin (BSA, Solarbio, China), Cell Counting Kit-8 (CCK-8, NCM, China), Bicin-choninic Acid (BCA, NCM, China), Enhanced chemiluminescent reagent (NCM, China), RIPA assay buffer (NCM, China), Blotting reagent (NCM, China), Color page gel rapid preparation kit (Epizyme, USA), Multicolor prestained protein ladder (Epizyme, USA), Mitotracker Red CMXRos (Beyotime, China), Mitochondrial membrane potential (ΔΨm) assay kit with JC-1 (Beyotime, China), ROS kit (Beyotime, China), 4′,6-Diamidino-2-phenylindole (DAPI, Beyotime, China), Hoechst (Beyotime, China), 4 % Paraformaldehyde (PFA, Solarbio, China), sodium citrate buffer (Solarbio, China), formalin-EDTA decalcifying solution (Solarbio, China), Hematoxylin-eosin stain kit (H&E, Solarbio, China), Modified Masson's trichrome stain kit (Masson, Solarbio, China), Triton X-100 (Servicebio, China), Enhanced endogenous peroxidase blocking buffer (Beyotime, China), Diaminobenzidine (DAB, Zsbio, China), Tris buffered saline with Tween-20 (TBST, Solarbio, China), TRIzol (Sigma, USA), ALN (Sigma, USA), Ex-4 (Selleck, USA), Dexamethasone (MCE, USA), β-Glycerophosphate disodium salt pentahydrate (MCE, USA) and Vitamin C (MCE, USA), Palmitic acid (Pa, Kunchuang Technology, China), Bafilomycin A1 (BafA1, MCE, USA).

Techniques: Inhibition, Expressing, Transfection

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 attenuate oxidative stress, preserve mitochondrial integrity, and suppress senescence in TSPCs by inhibiting the mtDNA-STING-NF-κB axis. (A) Schematic of the experimental design. (B) TSPC proliferation assessed by CCK-8 assay. (C, E) Immunofluorescence staining and quantification of HSP70 (n = 3). (D, F) Mitochondrial membrane potential (ΔΨm) visualized by JC-1 staining (red: high potential; green: low potential) and quantification. (G) Multi-SIM of mtDNA (magenta) and TOMM20 (green) with colocalization analysis. (H) Western blot of cGAS-STING-IRF3-NF-κB pathway proteins. (I, K) Colony-forming unit fibroblast (CFU-F) assay and quantification of self-renewal capacity (n = 3). (J) Senescence-associated β-galactosidase (SA-β-gal) activity. (L, M) Apoptosis analysis by Annexin V/PI flow cytometry and quantification (n = 3). Scale bars: 100 μm (C); 5 μm (D, G); 200 μm (J). Significance: ns, not significant; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: Mitochondrial membrane potential (ΔΨm) was evaluated via JC-1 staining (MedChemExpress, USA).

Techniques: CCK-8 Assay, Immunofluorescence, Staining, Membrane, Western Blot, Activity Assay, Flow Cytometry

Journal: International Journal of Nanomedicine

Article Title: Photothermolysis mediated by gold nanorods modified with EGFR monoclonal antibody induces Hep-2 cells apoptosis in vitro and in vivo

doi: 10.2147/IJN.S59870

Figure Lengend Snippet: ( A and B ) Effects of EGFRmAb-AuNRs+Laser treatment on caspase-3, Bax, ROS, Ca 2+ , ΔΨm, and Bcl-2 in Hep-2 cells. Note: Results represent one of three replicates. Abbreviations: ΔΨm, change in mitochondrial membrane potential; AuNRs, gold nanorods; Bax, Bcl-2-associated X protein; Bcl-2, B-cell lymphoma 2 protein; Cyt-c, cytochrome c; EGFRmAb, anti-EGFR monoclonal antibody; ROS, reactive oxygen species.

Article Snippet: Meanwhile, FCM was used to analyze other apoptotic molecules, including cytochrome c (Cyt-c) (CBA077 InnoCyteTM Flow Cytometric Cytochrome c Release Kit/CBA077-1KIT; EMD Millipore), change in mitochondrial membrane potential (ΔΨm) (Mitochondria staining kit (JC-1)/125T; MultiSciences Biotech Company, People’s Republic of China), B-cell lymphoma 2 protein (Bcl-2) (Bcl2 Mouse Anti-Human mAb [clone Bcl2/100]), FITC conjugate/A15764; Life Technologies), Bcl-2-associated X protein (Bax) (LifeSpan BioSciences Inc.), Ca 2+ (Calcium GreenTM-1/C-3012; Life Technologies), reactive oxygen species (ROS) (Reactive Oxygen Species Assay Kit/S0033; Beyotime Co, Jiangsu, People’s Republic of China) and caspase-3 (BD Biosciences, Franklin Lakes, NJ, USA).

Techniques: Membrane