caspase 12 inhibition  (R&D Systems)

Journal: International Journal of Medical Sciences

Article Title: Inhibiting Caspase-12 Mediated Inflammasome Activation protects against Oxygen-Glucose Deprivation Injury in Primary Astrocytes

doi: 10.7150/ijms.44330

Figure Lengend Snippet: NLRP3-inflammasome was expressed in primary mouse astrocytes. NLRP3-inflammasome components were existed in primary mouse astrocytes. ( A,B )The purity of primary mice astrocytes was detected by immunofluorescence and western blot (scale bar 20 µm, 400× magnification). ( C ) Immunofluorescence and was used to demonstrate the expression of NLRP3 and caspase-1 in primary mouse astrocytes in CON (control) group and OGD (3h hypoxia and 24h reperfusion) group (scale bar 20 µm, 600× or 1800x magnification). ( D ) Mean fluorescence intensity of NLRP3 and caspase-1 (comparisons to control group). Bar graphs were presented as means ± SD.*p<0.05, **p<0.01 (n=3).

Article Snippet: To verify if I/R-induced cell injury and apoptosis are mediated by caspase-12, primary mouse astrocytes were pretreated with the caspase-12 inhibitor Z-ATAD-FMK (ZAF; R&D; #FMK013).

Techniques: Immunofluorescence, Western Blot, Expressing, Control, Fluorescence

Journal: International Journal of Medical Sciences

Article Title: Inhibiting Caspase-12 Mediated Inflammasome Activation protects against Oxygen-Glucose Deprivation Injury in Primary Astrocytes

doi: 10.7150/ijms.44330

Figure Lengend Snippet: OGD induced cell injury and apoptosis in primary mouse astrocytes. Astrocytes were exposed to OGD for 1 to 4 hours and then returned to normal culture condition for 24 hours. ( A ) Cell injury was detected by LDH release assay. ( B ) Cell apoptosis was detected by Annexin V- fluorescein isothiocyanate (FITC)/prodium iodide (PI) flow cytometry, and the apoptosis rate was presented as the percentage of apoptotic cells vs total cells. ( C ) Representative scatterplots of Annexin V-FITC/PI flow cytometry in each group. CON: control group; OGD: Oxygen Glucose Deprivation group. * P < 0.05, ** P <0.01, *** P <0.001(all comparisons to control group). Bar graphs were presented as means ± SD. (n=6),

Article Snippet: To verify if I/R-induced cell injury and apoptosis are mediated by caspase-12, primary mouse astrocytes were pretreated with the caspase-12 inhibitor Z-ATAD-FMK (ZAF; R&D; #FMK013).

Techniques: Lactate Dehydrogenase Assay, Flow Cytometry, Control

Journal: International Journal of Medical Sciences

Article Title: Inhibiting Caspase-12 Mediated Inflammasome Activation protects against Oxygen-Glucose Deprivation Injury in Primary Astrocytes

doi: 10.7150/ijms.44330

Figure Lengend Snippet: OGD increased the expression of NLRP3 inflammasome components, including NLRP3 and caspase-1, as well as caspase-12 and IL-1β expression in primary mouse astrocytes. ( A ) Western blot analysis of NLRP3, caspase-1, IL-1β, caspase-12 and cleaved caspase-3 in astrocytes exposed to 1, 2, 3, 4 hours of OGD and then returned to normal condition for 24 hours. β-actin was used as a loading control. Protein levels of cleaved caspase-12 ( B ) NLRP3 ( C ), caspase-1 ( D ), IL-1β ( E ), and cleaved caspase-3 ( F ) were normalized to β-actin and quantified by Image Lab software. CON: control group; OGD: Oxygen Glucose Deprivation group. * P < 0.05, ** P <0.01, *** P <0.001(all comparisons to control group). Bar graphs were presented as means ± SD (n=6).

Article Snippet: To verify if I/R-induced cell injury and apoptosis are mediated by caspase-12, primary mouse astrocytes were pretreated with the caspase-12 inhibitor Z-ATAD-FMK (ZAF; R&D; #FMK013).

Techniques: Expressing, Western Blot, Control, Software

Journal: International Journal of Medical Sciences

Article Title: Inhibiting Caspase-12 Mediated Inflammasome Activation protects against Oxygen-Glucose Deprivation Injury in Primary Astrocytes

doi: 10.7150/ijms.44330

Figure Lengend Snippet: Inhibition of caspase-12 prevented OGD-induced cell injury in primary mouse astrocytes. Astrocytes were exposed to 3 hours of OGD and then 24 hours of reperfusion with or without ZAF (0, 0.05, 0.5, 5, 10 µM). ( A ) Cell injury was detected by LDH release assay. ( B ) Cell apoptosis was detected by Annexin V- fluorescein isothiocyanate (FITC)/prodium iodide (PI) flow cytometry, and the apoptosis rate was presented as the percentage of apoptotic cells vs total cells. ( C ) Representative scatterplots of Annexin V-FITC/PI flow cytometry in each group. CON: control group; OGD: Oxygen Glucose Deprivation group. * P < 0.05, ** P <0.01, *** P <0.001 (all comparisons to control (ZAF=0μM) group); # P < 0.05, ## P<0.01, ### P<0.001 (all comparisons to OGD(ZAF=0μM) group). Bar graphs were presented as means ± SD (n=6).

Article Snippet: To verify if I/R-induced cell injury and apoptosis are mediated by caspase-12, primary mouse astrocytes were pretreated with the caspase-12 inhibitor Z-ATAD-FMK (ZAF; R&D; #FMK013).

Techniques: Inhibition, Lactate Dehydrogenase Assay, Flow Cytometry, Control

Journal: International Journal of Medical Sciences

Article Title: Inhibiting Caspase-12 Mediated Inflammasome Activation protects against Oxygen-Glucose Deprivation Injury in Primary Astrocytes

doi: 10.7150/ijms.44330

Figure Lengend Snippet: Inhibition of caspase-12 attenuates NLRP3-inflammasome activation in OGD in primary mouse astrocytes. ( A ) Western blot analysis of NLRP3, caspase-1, IL-1β, caspase-12 and cleaved caspase-3 in astrocytes exposed to 3 hours of OGD followed by 24 hours reperfusion with or without ZAF (5 µM). β-actin was used as a loading control. Protein levels of cleaved caspase-12 ( B ) NLRP3 ( C ), caspase-1 ( D ), IL-1β ( E ), and cleaved caspase-3 ( F ) were normalized to β-actin and quantified by Image Lab software. CON: control group; OGD: Oxygen Glucose Deprivation group. * P < 0.05, ** P <0.01, *** P <0.001 (all comparisons to control (ZAF=0 µM) group); # P < 0.05, ## P<0.01, ### P<0.001 (all comparisons to OGD (ZAF=0 µM) group). Bar graphs were presented as means ± SD (n=6).

Article Snippet: To verify if I/R-induced cell injury and apoptosis are mediated by caspase-12, primary mouse astrocytes were pretreated with the caspase-12 inhibitor Z-ATAD-FMK (ZAF; R&D; #FMK013).

Techniques: Inhibition, Activation Assay, Western Blot, Control, Software