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Figure 6: Chloride intracellular channel-dependent chloride efflux induced pyroptosis mediated by NLRP3 inflammasomes in H9c2 cells. (a) Protein expression of CLIC1 and CLIC4 in heart tissues. (b) Relative mRNA expression of Clic1 and Clic4 in hearts. (c) IAA-94 was used to block chloride efflux, and the intracellular chloride was measured with MQAE, a chloride-sensitive dye. (d) Chloride concentrations in supernatants. (e) The LDH release measured by a <t>colorimetric</t> assay. (f) Caspase-1 activity. (g) E42 staining indicated the membrane integrity. (h) TUNEL staining of H9c2 cells. (i) Protein levels of NLRP3-related molecules. (j) The ratio of caspase-1 p12/procaspase-1 and (k) the ratio of phospho-NF-κB/total NF-κB. (l) Relative mRNA expression of Nlrp3-related molecules in H9c2 cells, Tubb3 as the internal reference. Data are presented as mean ± SD. ∗∗P < 0:01 CON vs. HG; ##P < 0:01 HG+IAA-94 vs. HG. n = 3 in each group.
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Figure 6: Chloride intracellular channel-dependent chloride efflux induced pyroptosis mediated by NLRP3 inflammasomes in H9c2 cells. (a) Protein expression of CLIC1 and CLIC4 in heart tissues. (b) Relative mRNA expression of Clic1 and Clic4 in hearts. (c) IAA-94 was used to block chloride efflux, and the intracellular chloride was measured with MQAE, a chloride-sensitive dye. (d) Chloride concentrations in supernatants. (e) The LDH release measured by a <t>colorimetric</t> assay. (f) Caspase-1 activity. (g) E42 staining indicated the membrane integrity. (h) TUNEL staining of H9c2 cells. (i) Protein levels of NLRP3-related molecules. (j) The ratio of caspase-1 p12/procaspase-1 and (k) the ratio of phospho-NF-κB/total NF-κB. (l) Relative mRNA expression of Nlrp3-related molecules in H9c2 cells, Tubb3 as the internal reference. Data are presented as mean ± SD. ∗∗P < 0:01 CON vs. HG; ##P < 0:01 HG+IAA-94 vs. HG. n = 3 in each group.
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Figure 6: Chloride intracellular channel-dependent chloride efflux induced pyroptosis mediated by NLRP3 inflammasomes in H9c2 cells. (a) Protein expression of CLIC1 and CLIC4 in heart tissues. (b) Relative mRNA expression of Clic1 and Clic4 in hearts. (c) IAA-94 was used to block chloride efflux, and the intracellular chloride was measured with MQAE, a chloride-sensitive dye. (d) Chloride concentrations in supernatants. (e) The LDH release measured by a colorimetric assay. (f) Caspase-1 activity. (g) E42 staining indicated the membrane integrity. (h) TUNEL staining of H9c2 cells. (i) Protein levels of NLRP3-related molecules. (j) The ratio of caspase-1 p12/procaspase-1 and (k) the ratio of phospho-NF-κB/total NF-κB. (l) Relative mRNA expression of Nlrp3-related molecules in H9c2 cells, Tubb3 as the internal reference. Data are presented as mean ± SD. ∗∗P < 0:01 CON vs. HG; ##P < 0:01 HG+IAA-94 vs. HG. n = 3 in each group.

Journal: Oxidative medicine and cellular longevity

Article Title: Attenuation of ROS/Chloride Efflux-Mediated NLRP3 Inflammasome Activation Contributes to Alleviation of Diabetic Cardiomyopathy in Rats after Sleeve Gastrectomy.

doi: 10.1155/2022/4608914

Figure Lengend Snippet: Figure 6: Chloride intracellular channel-dependent chloride efflux induced pyroptosis mediated by NLRP3 inflammasomes in H9c2 cells. (a) Protein expression of CLIC1 and CLIC4 in heart tissues. (b) Relative mRNA expression of Clic1 and Clic4 in hearts. (c) IAA-94 was used to block chloride efflux, and the intracellular chloride was measured with MQAE, a chloride-sensitive dye. (d) Chloride concentrations in supernatants. (e) The LDH release measured by a colorimetric assay. (f) Caspase-1 activity. (g) E42 staining indicated the membrane integrity. (h) TUNEL staining of H9c2 cells. (i) Protein levels of NLRP3-related molecules. (j) The ratio of caspase-1 p12/procaspase-1 and (k) the ratio of phospho-NF-κB/total NF-κB. (l) Relative mRNA expression of Nlrp3-related molecules in H9c2 cells, Tubb3 as the internal reference. Data are presented as mean ± SD. ∗∗P < 0:01 CON vs. HG; ##P < 0:01 HG+IAA-94 vs. HG. n = 3 in each group.

Article Snippet: The chloride concentration in the supernatants was examined using a Chloride Colorimetric Assay Kit (Elabscience, Wuhan, China).

Techniques: Expressing, Blocking Assay, Colorimetric Assay, Activity Assay, Staining, Membrane, TUNEL Assay