antibodies against cav 1  (Bioss)

Journal: Journal of Pharmaceutical Analysis

Article Title: Naringenin boosts Parkin-mediated mitophagy via estrogen receptor alpha to maintain mitochondrial quality control and heal diabetic foot ulcer

doi: 10.1016/j.jpha.2025.101333

Figure Lengend Snippet: Naringenin (NAR) accelerates wound healing in diabetic foot ulcer (DFU) mice. (A) Flowchart of DFU murine model construction and treatment. A round wound with a diameter of 6 mm was made on the back of each mouse. NAR (1%, 5%, 10%) or blank ointments (BOs) were applied around the wounds for 10 consecutive days. (B) The chemical structure of NAR. (C) The representative images of the wound healing process of mice at days 0, 1, 3, 5, 7, and 9 post-puncture ( n = 6). (D) Wound closure rates were calculated using images on days 1, 3, 5, 7, and 9 using ImageJ. (E) Representative images of histological assessment of gap closure in the wound epithelia of mice at day 9 post-puncture. (F, G) Representative immunohistochemistry (IHC) staining images of proliferation marker protein Ki67 and caveolin-1 (Cav-1) in wounds of mice at day 9 post-puncture. Data are presented as mean ± standard deviation (SD). ∗∗ P < 0.01 and ∗∗∗ P < 0.001, compared with the control group. HFD: high-fat diet; i.p.: intraperitoneal; STZ: streptomycin; H&E: hematoxylin and eosin.

Article Snippet: The slides were incubated with antibodies against caveolin-1 (Cav-1) (Sangon Biotech Co., Ltd.), proliferation marker protein Ki-67 (Abcam, Cambridge, UK), nicotinamide adenine dinucleotide phosphate (NAD(P)H): quinone oxidoreductase 1 (NQO1) (Sangon Biotech Co., Ltd.), and Parkin (Cell Signaling Technology, Danvers, MA, USA) at RT for 2 h. Afterward, they were incubated with secondary antibodies (Invitrogen, Carlsbad, CA, USA) at RT for 2 h. The samples were thereafter reacted with a 3,3′-diaminobenzidine (DAB) solution (Beijing Zsgb Bio, Beijing, China) to visualize signals, followed by counterstaining with hematoxylin.

Techniques: Immunohistochemistry, Marker, Standard Deviation, Control

Journal: Journal of Pharmaceutical Analysis

Article Title: Naringenin boosts Parkin-mediated mitophagy via estrogen receptor alpha to maintain mitochondrial quality control and heal diabetic foot ulcer

doi: 10.1016/j.jpha.2025.101333

Figure Lengend Snippet: Naringenin (NAR) alleviates high glucose (HG)-induced keratinocyte dysfunction. HaCaT cells were treated with NAR (0.12, 0.37, 1.1, and 3.3 μM) for 24 h after being induced with HG (50 mM). (A) Immunofluorescence staining was used to detect the proliferation marker protein Ki67 expression in HaCaT cells. (B, C) The senescence of HaCaT cells was detected by senescence-associated β-galactosidase (SA-β-gal) staining (B) and quantification analysis (C). (D) The expression levels of senescence-related proteins, including LaminB1, cyclin-dependent kinase inhibitor 1A (P21), caveolin-1 (Cav-1), and phospho-H2A histone family member X (γH2AX) in HaCaT cells were measured by Western blotting. (E) The protein levels of nuclear factor-κB (NF-κB) and -NF-κB (S536) in HaCaT cells were measured by Western blotting. (F, G) The apoptotic rates of HaCaT cells were determined by flow cytometry (F) and quantification analysis (G). Two-way analysis of variance was performed for statistical analysis. (H) Representative images of the cell scratch assay in HaCaT cells. (I) The wound closure rate was calculated by measuring the scratch area using ImageJ. Data are presented as mean ± standard deviation (SD). ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001, compared with the control group. GAPDH: glyceraldehyde 3-phosphate dehydrogenase; PI: propidium iodide; FITC: fluorescein isothiocyanate.

Article Snippet: The slides were incubated with antibodies against caveolin-1 (Cav-1) (Sangon Biotech Co., Ltd.), proliferation marker protein Ki-67 (Abcam, Cambridge, UK), nicotinamide adenine dinucleotide phosphate (NAD(P)H): quinone oxidoreductase 1 (NQO1) (Sangon Biotech Co., Ltd.), and Parkin (Cell Signaling Technology, Danvers, MA, USA) at RT for 2 h. Afterward, they were incubated with secondary antibodies (Invitrogen, Carlsbad, CA, USA) at RT for 2 h. The samples were thereafter reacted with a 3,3′-diaminobenzidine (DAB) solution (Beijing Zsgb Bio, Beijing, China) to visualize signals, followed by counterstaining with hematoxylin.

Techniques: Immunofluorescence, Staining, Marker, Expressing, Western Blot, Flow Cytometry, Wound Healing Assay, Standard Deviation, Control

Journal: Journal of Pharmaceutical Analysis

Article Title: Naringenin boosts Parkin-mediated mitophagy via estrogen receptor alpha to maintain mitochondrial quality control and heal diabetic foot ulcer

doi: 10.1016/j.jpha.2025.101333

Figure Lengend Snippet: Naringenin (NAR)'s effects on diabetic wound healing are abolished in E3 ubiquitin-protein ligase parkin (Parkin/PRKN/Prkn) deficiency mice. (A) Flowchart of Prkn knockout ( Prkn − /− ) diabetic foot ulcer (DFU) murine model construction and treatment. The four-week-old male Prkn − /− mice were utilized to confirm the role of Parkin in NAR's effects on DFU. After genotyping, the mice were intraperitoneally injected with streptozotocin (STZ) at a dose of 50 mg/kg for five consecutive days. Circular wounds of 6-mm diameter on the back of mice with fasting blood glucose (FBG) level above 200 mg/dL were created by a skin sampler. Blank or 5% NAR ointment was applied around the wounds for 10 consecutive days after injury. Each group of mice was maintained on a normal chow diet (CD) throughout the experiment. (B) Representative images of the skin wounds of Prkn − /− mice on days 0, 1, 3, 5, 7, and 9 post-puncture ( n = 5). (C) Wound closure rates in B were calculated using images captured on days 1, 3, 5, 7, and 9 through ImageJ. (D) Representative images of histological assessment of gap closure in the wound epithelia of Prkn − /− mice on day 9 post-puncture. (E–H) Representative immunohistochemistry (IHC) staining images of Parkin (E), proliferation marker protein Ki67 (F), caveolin-1 (Cav-1) (G), and nicotinamide adenine dinucleotide phosphate (NAD(P)H):quinone oxidoreductase 1 (NQO1) (H) in the wound of Prkn − /− mice on day 9 post-puncture. Data are presented as mean ± standard deviation (SD). ∗ P < 0.05, compared with the control group. ND: normal diet; i.p.: intraperitoneal; H&E: hematoxylin and eosin.

Article Snippet: The slides were incubated with antibodies against caveolin-1 (Cav-1) (Sangon Biotech Co., Ltd.), proliferation marker protein Ki-67 (Abcam, Cambridge, UK), nicotinamide adenine dinucleotide phosphate (NAD(P)H): quinone oxidoreductase 1 (NQO1) (Sangon Biotech Co., Ltd.), and Parkin (Cell Signaling Technology, Danvers, MA, USA) at RT for 2 h. Afterward, they were incubated with secondary antibodies (Invitrogen, Carlsbad, CA, USA) at RT for 2 h. The samples were thereafter reacted with a 3,3′-diaminobenzidine (DAB) solution (Beijing Zsgb Bio, Beijing, China) to visualize signals, followed by counterstaining with hematoxylin.

Techniques: Ubiquitin Proteomics, Knock-Out, Injection, Ointment, Immunohistochemistry, Marker, Standard Deviation, Control