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USP18 aggravates cardiac I/R injury through regulation of mitochondria and inhibition of mitophagy. a Electron microscopy image showing mitophagy in USP18-cKO mouse hearts ( n= 5). Scale bar=3 μm. White arrowheads indicate sites of mitophagy. b Protein levels of PINK1, Parkin, ubiquitinated proteins (Ub), P62, and LC3II in mitochondria from heart tissue in USP18-cKO and WT mice 24 h after I/R injury ( n =4). c Electron microscopy image showing mitophagy in USP18-overexpres (OV) mouse hearts ( n =5). Scale bar=3 μm. White arrowheads indicate sites of mitophagy. d Protein levels of PINK1, Parkin, Ub, P62, and LC3II proteins in mitochondria from the heart tissue of USP18-OV mice 24 h after I/R injury ( n =4). Color shift in mitophagy dye (red) and lysosomal dye (green) in NRVMs showing mitophagy in NRVMs with USP18 siRNA transfection ( e ) or Ad-USP18 infection ( f ) and the quantitative mitophagy index in each group ( n= 5). Scale bar=9 μm. Protein levels of PINK1, Parkin, Ub, P62, and LC3II in mitochondria from NRVMs transfected with USP18 siRNA ( g ) or infected with Ad-USP18 ( h ). ⁎⁎ P <0.01, ⁎⁎⁎ P <0.001 ⁎⁎⁎⁎ P <0.0001. USP18. Ubiquitin-specific protease 18; I/R. Ischemia/reperfusion; WT. Wild-type; KO. Knockout; P62. Sequestosome 1; LC3. Microtubule-associated <t>protein</t> <t>1</t> light chain 3; VDAC. Voltage-dependent anion channel.
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USP18 aggravates cardiac I/R injury through regulation of mitochondria and inhibition of mitophagy. a Electron microscopy image showing mitophagy in USP18-cKO mouse hearts ( n= 5). Scale bar=3 μm. White arrowheads indicate sites of mitophagy. b Protein levels of PINK1, Parkin, ubiquitinated proteins (Ub), P62, and LC3II in mitochondria from heart tissue in USP18-cKO and WT mice 24 h after I/R injury ( n =4). c Electron microscopy image showing mitophagy in USP18-overexpres (OV) mouse hearts ( n =5). Scale bar=3 μm. White arrowheads indicate sites of mitophagy. d Protein levels of PINK1, Parkin, Ub, P62, and LC3II proteins in mitochondria from the heart tissue of USP18-OV mice 24 h after I/R injury ( n =4). Color shift in mitophagy dye (red) and lysosomal dye (green) in NRVMs showing mitophagy in NRVMs with USP18 siRNA transfection ( e ) or Ad-USP18 infection ( f ) and the quantitative mitophagy index in each group ( n= 5). Scale bar=9 μm. Protein levels of PINK1, Parkin, Ub, P62, and LC3II in mitochondria from NRVMs transfected with USP18 siRNA ( g ) or infected with Ad-USP18 ( h ). ⁎⁎ P <0.01, ⁎⁎⁎ P <0.001 ⁎⁎⁎⁎ P <0.0001. USP18. Ubiquitin-specific protease 18; I/R. Ischemia/reperfusion; WT. Wild-type; KO. Knockout; P62. Sequestosome 1; LC3. Microtubule-associated <t>protein</t> <t>1</t> light chain 3; VDAC. Voltage-dependent anion channel.
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OBNC microspheres activate integrin receptors and mechanosensitive calcium channels. A) Mechanistic diagram of integrin activation verified using fluorophores. B-C) Fluorescence microscopy images of MSCs loaded on HAMA or OBNC hydrogel. D) Fluorescence intensity of single cell in each group was quantified. E) Fluorescence microscopy of MSCs loaded on OBNC hydrogel after different treatments. F) Fluorescence intensity in the whole field of view for each group. G) Fluorescence intensity in the single cell for each group. H) Schematic representation of patch clamp experiments. I) Electrical signals generated by MSCs in response to mechanical stimulation. J) Statistical analysis of poking currents (n = 6). K) The concentration of calcium ions in stem cells of different groups as detected by flow cytometry (siRNA1: targeting the <t>TRPM4</t> gene, siRNA2: targeting the PIEZO1 gene). L) Quantitative analysis of flow cytometric results (∗ symbol represents comparison with HAMA group, # symbol represents comparison with OBNC group). (ns: non-significant, ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ## P < 0.01, ### P < 0.001).
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OBNC microspheres activate integrin receptors and mechanosensitive calcium channels. A) Mechanistic diagram of integrin activation verified using fluorophores. B-C) Fluorescence microscopy images of MSCs loaded on HAMA or OBNC hydrogel. D) Fluorescence intensity of single cell in each group was quantified. E) Fluorescence microscopy of MSCs loaded on OBNC hydrogel after different treatments. F) Fluorescence intensity in the whole field of view for each group. G) Fluorescence intensity in the single cell for each group. H) Schematic representation of patch clamp experiments. I) Electrical signals generated by MSCs in response to mechanical stimulation. J) Statistical analysis of poking currents (n = 6). K) The concentration of calcium ions in stem cells of different groups as detected by flow cytometry (siRNA1: targeting the <t>TRPM4</t> gene, siRNA2: targeting the PIEZO1 gene). L) Quantitative analysis of flow cytometric results (∗ symbol represents comparison with HAMA group, # symbol represents comparison with OBNC group). (ns: non-significant, ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ## P < 0.01, ### P < 0.001).
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OBNC microspheres activate integrin receptors and mechanosensitive calcium channels. A) Mechanistic diagram of integrin activation verified using fluorophores. B-C) Fluorescence microscopy images of MSCs loaded on HAMA or OBNC hydrogel. D) Fluorescence intensity of single cell in each group was quantified. E) Fluorescence microscopy of MSCs loaded on OBNC hydrogel after different treatments. F) Fluorescence intensity in the whole field of view for each group. G) Fluorescence intensity in the single cell for each group. H) Schematic representation of patch clamp experiments. I) Electrical signals generated by MSCs in response to mechanical stimulation. J) Statistical analysis of poking currents (n = 6). K) The concentration of calcium ions in stem cells of different groups as detected by flow cytometry (siRNA1: targeting the <t>TRPM4</t> gene, siRNA2: targeting the PIEZO1 gene). L) Quantitative analysis of flow cytometric results (∗ symbol represents comparison with HAMA group, # symbol represents comparison with OBNC group). (ns: non-significant, ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ## P < 0.01, ### P < 0.001).
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OBNC microspheres activate integrin receptors and mechanosensitive calcium channels. A) Mechanistic diagram of integrin activation verified using fluorophores. B-C) Fluorescence microscopy images of MSCs loaded on HAMA or OBNC hydrogel. D) Fluorescence intensity of single cell in each group was quantified. E) Fluorescence microscopy of MSCs loaded on OBNC hydrogel after different treatments. F) Fluorescence intensity in the whole field of view for each group. G) Fluorescence intensity in the single cell for each group. H) Schematic representation of patch clamp experiments. I) Electrical signals generated by MSCs in response to mechanical stimulation. J) Statistical analysis of poking currents (n = 6). K) The concentration of calcium ions in stem cells of different groups as detected by flow cytometry (siRNA1: targeting the <t>TRPM4</t> gene, siRNA2: targeting the PIEZO1 gene). L) Quantitative analysis of flow cytometric results (∗ symbol represents comparison with HAMA group, # symbol represents comparison with OBNC group). (ns: non-significant, ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ## P < 0.01, ### P < 0.001).
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Image Search Results


USP18 aggravates cardiac I/R injury through regulation of mitochondria and inhibition of mitophagy. a Electron microscopy image showing mitophagy in USP18-cKO mouse hearts ( n= 5). Scale bar=3 μm. White arrowheads indicate sites of mitophagy. b Protein levels of PINK1, Parkin, ubiquitinated proteins (Ub), P62, and LC3II in mitochondria from heart tissue in USP18-cKO and WT mice 24 h after I/R injury ( n =4). c Electron microscopy image showing mitophagy in USP18-overexpres (OV) mouse hearts ( n =5). Scale bar=3 μm. White arrowheads indicate sites of mitophagy. d Protein levels of PINK1, Parkin, Ub, P62, and LC3II proteins in mitochondria from the heart tissue of USP18-OV mice 24 h after I/R injury ( n =4). Color shift in mitophagy dye (red) and lysosomal dye (green) in NRVMs showing mitophagy in NRVMs with USP18 siRNA transfection ( e ) or Ad-USP18 infection ( f ) and the quantitative mitophagy index in each group ( n= 5). Scale bar=9 μm. Protein levels of PINK1, Parkin, Ub, P62, and LC3II in mitochondria from NRVMs transfected with USP18 siRNA ( g ) or infected with Ad-USP18 ( h ). ⁎⁎ P <0.01, ⁎⁎⁎ P <0.001 ⁎⁎⁎⁎ P <0.0001. USP18. Ubiquitin-specific protease 18; I/R. Ischemia/reperfusion; WT. Wild-type; KO. Knockout; P62. Sequestosome 1; LC3. Microtubule-associated protein 1 light chain 3; VDAC. Voltage-dependent anion channel.

Journal: Military Medical Research

Article Title: USP18 exacerbates myocardial I/R injury by inhibiting Parkin mitophagy through the deubiquitinase PTEN-L

doi: 10.1016/j.mmr.2026.100004

Figure Lengend Snippet: USP18 aggravates cardiac I/R injury through regulation of mitochondria and inhibition of mitophagy. a Electron microscopy image showing mitophagy in USP18-cKO mouse hearts ( n= 5). Scale bar=3 μm. White arrowheads indicate sites of mitophagy. b Protein levels of PINK1, Parkin, ubiquitinated proteins (Ub), P62, and LC3II in mitochondria from heart tissue in USP18-cKO and WT mice 24 h after I/R injury ( n =4). c Electron microscopy image showing mitophagy in USP18-overexpres (OV) mouse hearts ( n =5). Scale bar=3 μm. White arrowheads indicate sites of mitophagy. d Protein levels of PINK1, Parkin, Ub, P62, and LC3II proteins in mitochondria from the heart tissue of USP18-OV mice 24 h after I/R injury ( n =4). Color shift in mitophagy dye (red) and lysosomal dye (green) in NRVMs showing mitophagy in NRVMs with USP18 siRNA transfection ( e ) or Ad-USP18 infection ( f ) and the quantitative mitophagy index in each group ( n= 5). Scale bar=9 μm. Protein levels of PINK1, Parkin, Ub, P62, and LC3II in mitochondria from NRVMs transfected with USP18 siRNA ( g ) or infected with Ad-USP18 ( h ). ⁎⁎ P <0.01, ⁎⁎⁎ P <0.001 ⁎⁎⁎⁎ P <0.0001. USP18. Ubiquitin-specific protease 18; I/R. Ischemia/reperfusion; WT. Wild-type; KO. Knockout; P62. Sequestosome 1; LC3. Microtubule-associated protein 1 light chain 3; VDAC. Voltage-dependent anion channel.

Article Snippet: To block mitophagy, the selective dynamin-related protein 1 (Drp1) inhibitor Mdivi-1 was used (50 μmol/L, MedChemExpress, USA).

Techniques: Inhibition, Electron Microscopy, Transfection, Infection, Ubiquitin Proteomics, Knock-Out

OBNC microspheres activate integrin receptors and mechanosensitive calcium channels. A) Mechanistic diagram of integrin activation verified using fluorophores. B-C) Fluorescence microscopy images of MSCs loaded on HAMA or OBNC hydrogel. D) Fluorescence intensity of single cell in each group was quantified. E) Fluorescence microscopy of MSCs loaded on OBNC hydrogel after different treatments. F) Fluorescence intensity in the whole field of view for each group. G) Fluorescence intensity in the single cell for each group. H) Schematic representation of patch clamp experiments. I) Electrical signals generated by MSCs in response to mechanical stimulation. J) Statistical analysis of poking currents (n = 6). K) The concentration of calcium ions in stem cells of different groups as detected by flow cytometry (siRNA1: targeting the TRPM4 gene, siRNA2: targeting the PIEZO1 gene). L) Quantitative analysis of flow cytometric results (∗ symbol represents comparison with HAMA group, # symbol represents comparison with OBNC group). (ns: non-significant, ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ## P < 0.01, ### P < 0.001).

Journal: Bioactive Materials

Article Title: Mechanically sensitized hydrogel microspheres trigger membrane receptor switch for cartilage repair

doi: 10.1016/j.bioactmat.2026.03.017

Figure Lengend Snippet: OBNC microspheres activate integrin receptors and mechanosensitive calcium channels. A) Mechanistic diagram of integrin activation verified using fluorophores. B-C) Fluorescence microscopy images of MSCs loaded on HAMA or OBNC hydrogel. D) Fluorescence intensity of single cell in each group was quantified. E) Fluorescence microscopy of MSCs loaded on OBNC hydrogel after different treatments. F) Fluorescence intensity in the whole field of view for each group. G) Fluorescence intensity in the single cell for each group. H) Schematic representation of patch clamp experiments. I) Electrical signals generated by MSCs in response to mechanical stimulation. J) Statistical analysis of poking currents (n = 6). K) The concentration of calcium ions in stem cells of different groups as detected by flow cytometry (siRNA1: targeting the TRPM4 gene, siRNA2: targeting the PIEZO1 gene). L) Quantitative analysis of flow cytometric results (∗ symbol represents comparison with HAMA group, # symbol represents comparison with OBNC group). (ns: non-significant, ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ## P < 0.01, ### P < 0.001).

Article Snippet: TRPC1 inhibitor (0.3 nM, Pico145, CAS No. 1628287-16-0), TRPM7 inhibitor (1.0 μM, VPC4, CAS No. 945604-76-2), TRPV2 inhibitor (5.0 μM, compound IV2-1, CAS No. 2242724-49-6), TRPM4 inhibitor (1.5 μM, CBA, CAS No. 351424-20-9), PIEZO1 inhibitor (2.5 μM, GsMTx4, CAS No. 1209500-46-8), integrin αvβ5 inhibitor (8.0 nM, Compound 12, CAS No.: 2615912-33-7), integrin αvβ1 inhibitor (0.3 nM, Compound C8, CAS No. 1689540-62-2), integrin α5β1 inhibitor (10 μM, ATN-161, 904763-27-5), and CDK5 inhibitor (5 nM, CDK5-IN-1, 2,639,540-19-3) were purchased from MCE Biotechnology Co., LTD. After the MSCs were treated, the cRGD solution was added at a concentration of 1:200 and incubated in the dark for 15 min, and the results were observed by fluorescence microscopy.

Techniques: Activation Assay, Fluorescence, Microscopy, Single Cell, Patch Clamp, Generated, Concentration Assay, Flow Cytometry, Comparison