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Image Search Results
Journal: Advanced Science
Article Title: A Bioinspired Manganese‐Organic Framework Ameliorates Ischemic Stroke through its Intrinsic Nanozyme Activity and Upregulating Endogenous Antioxidant Enzymes
doi: 10.1002/advs.202206854
Figure Lengend Snippet: The effect of pDA‐MNOF on upregulating HO1 and SOD2 via STAT3 signaling. a) Western blot analysis of p‐STAT3, STAT3, HO1, and SOD2 expression in N2a cells treated with 12.5 µg mL −1 pDA‐MNOF for various time durations. b) Quantification of the band intensity ratios of p‐STAT3/STAT3, HO1/ β ‐actin, and SOD2/ β ‐actin in (a). t‐STAT3 indicated total proteins of p‐STAT3 and STAT3. c) Quantification of fluorescent intensity of p‐STAT3, HO1, and SOD2 in PBS or 12.5 µg mL −1 pDA‐MNOF‐treated N2a cells ( n = 4). Data were presented with mean ± s.d.; *, p < 0.05; **, p < 0.01; T ‐test. d) Representative images of immunofluorescent staining for p‐STAT3, HO‐1, and SOD‐2 (red) in indicated groups. Nuclei were stained in blue, and cell body was outlined with white dashed lines. Scale bar, 20 µm. e) Western blot analysis of p‐STAT3, STAT3, HO1, and SOD2 expression in N2a cells treated with different concentrations of pDA‐MNOF for 12 h. f) Quantification of the band intensity ratios of p‐STAT3/STAT3, HO1/ β ‐actin, and SOD2/ β ‐actin in (e). g) Experimental scheme for the RNA interference and pathway inhibition assays. h) Western blot analysis of p‐STAT3 and STAT3 in N2a cells treated with PBS, siRNA Control (siRNA Ctrl), siRNA 1#, siRNA 2#, and siRNA 3#. i) Quantification of the band intensity ratios of p‐STAT3/ β ‐actin and STAT3/ β ‐actin in (h). j) N2a cells were transfected with siRNA Ctrl, siRNA 2#, and siRNA 3#, followed by treatment with pDA‐MNOF; the lysates were analyzed with indicated antibodies. k) Quantification of the band intensity ratios of p‐STAT3/ β ‐actin, HO1/ β ‐actin, and SOD2/ β ‐actin in (j). Data were presented with mean ± s.d.; *, p < 0.05; ANOVA. l) N2a cells were treated with PBS or pDA‐MNOF, followed by treatment with AG490; the lysates were analyzed with indicated antibodies. m) Quantification of the band intensity ratios of p‐STAT3/STAT3, HO1/ β ‐actin, and SOD2/ β ‐actin in (l) ( n = 3). Data were presented with mean ± s.d.; *, p < 0.05; **, p < 0.01; ANOVA.
Article Snippet: To assess the protein levels of VEGF, the supernatant of
Techniques: Western Blot, Expressing, Staining, Inhibition, Control, Transfection
Journal: Advanced Science
Article Title: A Bioinspired Manganese‐Organic Framework Ameliorates Ischemic Stroke through its Intrinsic Nanozyme Activity and Upregulating Endogenous Antioxidant Enzymes
doi: 10.1002/advs.202206854
Figure Lengend Snippet: pDA‐MNOF protects neuronal cells against ROS‐induced injury. a) DCFH‐DA staining for detecting ROS in the PBS or pDA‐MNOF‐treated N2a cells after being treated with an 8 h OGD. N2a cells cultured in regular condition were set as the control. Scale bars, 50 µm. b) Quantification of percentages of DCFH‐DA positive (DCFDA + ) cells in indicated groups in (a) ( n = 5). Data were presented with mean ± s.d.; *, p < 0.05; ****, p < 0.0001; ANOVA. c) Flow cytometric analysis of DCFH‐DA positive cell proportions in the three groups. d) Fluorescence intensity detection in cell lysis of DCFH‐DA‐stained N2a cells in indicated groups ( n = 5). Data were presented with mean ± s.d.; *, p < 0.05; ***, p < 0.001; ANOVA. e) Schematic illustration of determining the contribution ratios of two effects of pDA‐MNOF on scavenging ROS. Briefly, N2a cells were treated with PBS, or 12.5 µg mL −1 pDA‐MNOF for 12 h, and their SOD‐like activities were determined as A 0 and A 1 , respectively. Cell lysis of pDA‐MNOF‐treated N2a cells was subject to ultracentrifugation for removing the residual pDA‐MNOF, and the SOD‐like activity was determined as A 3 . f) Quantification of contribution ratios of two effects of pDA‐MNOF on scavenging ROS. The formula was listed to the right. g) Live & Dead staining for PBS or pDA‐MNOF‐treated N2a cells after an 8 h OGD followed by a 16 h regular incubation. Scale bars, 100 µm. h) Quantification of percentages of dying cells in indicated groups in (g) ( n = 5). Data were presented with mean ± s.d.; ****, p < 0.0001; ANOVA. i) Cell viability of N2a cells in indicated groups in CCK‐8 assay ( n = 5). Data were presented with mean ± s.d.; **, p < 0.01; ****, p < 0.0001; ANOVA. j) Schematics of pDA‐MNOF utilizing nanozyme activity and upregulating endogenous HO1/SOD2, jointly scavenging ROS.
Article Snippet: To assess the protein levels of VEGF, the supernatant of
Techniques: Staining, Cell Culture, Control, Fluorescence, Lysis, Activity Assay, Incubation, CCK-8 Assay
Journal: Advanced Science
Article Title: A Bioinspired Manganese‐Organic Framework Ameliorates Ischemic Stroke through its Intrinsic Nanozyme Activity and Upregulating Endogenous Antioxidant Enzymes
doi: 10.1002/advs.202206854
Figure Lengend Snippet: pDA‐MNOF promotes angiogenesis. a) Schematics showing VEGF, a downstream gene of STAT3 signaling, was activated by pDA‐MNOF. b) The mRNA levels of VEGF in N2a cells treated with PBS (control) or pDA‐MNOF ( n = 3). Data were presented with mean ± s.d.; **, p < 0.01; ****, p < 0.0001; ANOVA. c) The levels of VEGF protein secreted from the PBS (control) or pDA‐MNOF‐treated N2a cells ( n = 3). Data were presented with mean ± s.d.; ***, p < 0.001; ****, p < 0.0001; ANOVA. d) Capillary‐like tube formation assay performed on C166 cells (murine endothelial cell line) that were cultured with regular N2a culture medium or conditioned medium obtained from the pDA‐MNOF‐treated N2a cells. 5pDA‐MNOF, 12.5pDA‐MNOF, or 25pDA‐MNOF represent conditioned medium that was from the 5, 12.5, or 25 µg mL −1 pDA‐MNOF‐treated N2a cells, respectively. The branch points were indicated by red arrowheads, and the structure between two adjacent arrowheads was a branch. e) Quantification of branch point numbers, tube length, and tube numbers in capillary‐like tube formation assay in (d) ( n = 5 repeats). f) Schematics of cerebral ventricle injection of PBS and pDA‐MNOF for further evaluating in vivo angiogenesis. g) Relative VEGF mRNA levels in brain tissue of the MCAo mice receiving cerebral ventricle injection of PBS or pDA‐MNOF ( n = 9). The mice received sham operation were set as control ( n = 3). Data were presented with mean ± s.d.; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001; ANOVA. h) Representative fluorescence images of vessels (CD31, red) and astrocytes (GFAP, green) in the infarct (an asterisk) and peri‐infarct 4 weeks after the mice received cerebral ventricle injection of PBS or pDA‐MNOF. The boundary between the infarct and peri‐infarct was indicated by white dashed lines. i,j) Quantification of percentages of CD31 positive (CD31 + ) area in i) the infarct and j) peri‐infarct, respectively ( n = 9). Data were presented with mean ± s.d.; *, p < 0.05; **, p < 0.01; ANOVA.
Article Snippet: To assess the protein levels of VEGF, the supernatant of
Techniques: Control, Capillary Tube Formation Assay, Cell Culture, Injection, In Vivo, Fluorescence