Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: VAMP4 in hypoxic adipose stem cell exosomes alleviates ischemia-reperfusion injury

doi: 10.1007/s00018-025-05967-4

Figure Lengend Snippet: Hypoxic ADSC-Ex alleviated oxidative stress in OGD/R-treated HUVECs A . Representative curves of ROS fluorescence detected via DCF staining in flow cytometry across different groups. B . Flow cytometric analysis of ROS ( n = 6). Hypoxic ADSC-Ex significantly reduced ROS levels in OGD/R-treated HUVECs. C . Western blotting analysis for Nox4, Nox1, NQO1, and SOD2 in OGD/R-treated control HUVECs, OGD/R HUVECs treated with ADSC-Ex, and OGD/R HUVECs treated with hypoxic ADSC-Ex. Nox4 and Nox1 levels decreased in the hypoxic ADSC-Ex group, while NQO1 and SOD2 levels increased in this group. D . Representative JC-1 staining images of HUVECs in different groups (Scale bar = 100 μm). E . Flow cytometric analysis of JC-1 staining in HUVECs across different groups ( n = 6). F . Statistical analysis of the red/green ratio from JC-1 staining in HUVECs across different groups. HUVECs treated with hypoxic ADSC-Ex showed a significantly increased red/green ratio. **** P < 0.0001.Abbreviations: ADSC-Ex, ADSC-derived exosomes; OGD/R, oxygen-glucose deprivation/reperfusion; NQO1, NAD(P)H: quinone oxidoreductase 1; Nox1, NADPH oxidase 1; Nox4, NADPH oxidase 4; SOD2, superoxide dismutase 2; ROS, reactive oxygen species; DCF, dichlorofluorescein; HUVECs, human umbilical vein endothelial cells; ADSC, adipose-derived stem cell

Article Snippet: For muscle tissue staining, sections were incubated overnight at 4°Cwith primary antibodies against CD31 (Abcam, #182981, 1:500), NQO1 (Affinity, #DF6437, 1:100), and SOD2 (Proteintech, #66474-1-Ig, 1:200).

Techniques: Fluorescence, Staining, Flow Cytometry, Western Blot, Control, Derivative Assay

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: VAMP4 in hypoxic adipose stem cell exosomes alleviates ischemia-reperfusion injury

doi: 10.1007/s00018-025-05967-4

Figure Lengend Snippet: Histological changes in the hypoxic ADSC-Ex-treated hind-limb I/RI model A . Histological injury score of skeletal muscle sections stained with Hematoxylin and Eosin ( n = 6). B . Representative images of skeletal muscle sections stained with antibodies against CD31 (red), TUNEL (green), and DAPI (blue) (Scale bar = 50 μm). C . Fluorescence intensity of CD31 in sham, I/RI control, and I/RI hypoxic ADSC-Ex treated groups ( n = 6). D . Quantification of TUNEL-positive cells. E . Quantification of NQO1 intensity within CD31 + cells. F . Quantification of SOD2 intensity within CD31 + cells. Data are expressed as means ± standard deviation (SD) ( n = 6). * P < 0.05,** P < 0.01, *** P < 0.001. Abbreviations: ADSC-Ex, ADSC-derived exosomes; I/RI, ischemia/reperfusion injury; CD31, cluster of differentiation 31; DAPI, 4’,6-diamidino-2-phenylindole

Article Snippet: For muscle tissue staining, sections were incubated overnight at 4°Cwith primary antibodies against CD31 (Abcam, #182981, 1:500), NQO1 (Affinity, #DF6437, 1:100), and SOD2 (Proteintech, #66474-1-Ig, 1:200).

Techniques: Staining, TUNEL Assay, Fluorescence, Control, Standard Deviation, Derivative Assay

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: VAMP4 in hypoxic adipose stem cell exosomes alleviates ischemia-reperfusion injury

doi: 10.1007/s00018-025-05967-4

Figure Lengend Snippet: The antioxidative effect of VAMP4 in hypoxic ADSC-Ex and its association with SNAP23 and NF-κB signaling A . Western blotting analysis for Nox4, Nox1, NQO1, and SOD2 in OGD/R-treated HUVECs treated with hypoxic ADSC-Ex, hypoxic ADSC-Ex (treated with si-NC), or hypoxic ADSC-Ex (treated with si-VAMP4). Nox4 and Nox1 levels increased in the si-VAMP4 group, while NQO1 and SOD2 levels decreased in the si-VAMP4 group. B . Representative flow cytometric analysis of JC-1 staining in OGD/R HUVECs across different groups. C . Statistical analysis of the red/green ratio from JC-1 staining in OGD/R HUVECs across different groups ( n = 6). Knockdown of VAMP4 in hypoxic ADSC-Ex significantly decreased the red/green ratio in OGD/R HUVECs treated with hypoxic ADSC-Ex. D . Western blotting analysis for Stx16, Stx6 and SNAP23 of OGD/R treated HUVECs in control, hypoxic ADSC-Ex, and hypoxic ADSC-Ex (treated with si-VAMP4) groups. E . Western blot analysis was conducted to evaluate IKKβ, p-P65, P65, and IKBα levels in cell lysates of OGD/R-treated HUVECs from the control, hypoxic ADSC-Ex, and hypoxic ADSC-Ex (treated with si-VAMP4) groups. F . Western blot analysis was conducted to evaluate p-P65 and P65 levels in nuclear proteins of OGD/R-treated HUVECs from the control, hypoxic ADSC-Ex, and hypoxic ADSC-Ex (treated with si-VAMP4) groups. ** P <0.01. Abbreviations: ADSC-Ex, ADSC-derived exosomes; HUVECs, human umbilical vein endothelial cells; ADSC, adipose-derived stem cell; OGD/R, oxygen-glucose deprivation; NQO1, NAD(P)H: quinone oxidoreductase 1; Nox1, NADPH oxidase 1; Nox4, NADPH oxidase 4; SOD2, Superoxide Dismutase 2; SNAP23, Synaptosomal-associated protein 23; Stx16, Syntaxin 16; Stx6, Syntaxin 6

Article Snippet: For muscle tissue staining, sections were incubated overnight at 4°Cwith primary antibodies against CD31 (Abcam, #182981, 1:500), NQO1 (Affinity, #DF6437, 1:100), and SOD2 (Proteintech, #66474-1-Ig, 1:200).

Techniques: Western Blot, Staining, Knockdown, Control, Derivative Assay