Figure 6.
AAV-mediated restore of CTCF expression ameliorates β cell dysfunction ex vivo and in vivo. (a) Schematic representation of experimental procedures for b–d. (b) Immunoblots of total cell lysates from mouse or human islets transduced with AAVs expressing CTCF or GFP for 5 d, followed by treatment with TNFα (50 ng/ml) and PA (0.5 mM) for an additional 24 h. (c) Perifusion analyses of dynamic glucose-stimulated insulin secretion (left) and biphasic insulin release levels (right) of mouse islets as described in b. Each islet sample was pooled from at least three animals. Data represent mean ± SD; n = 3 technical replicates; ***, P < 0.001; two-tailed unpaired Student’s t test. (d) Perifusion analyses of dynamic glucose-stimulated insulin secretion (left) and biphasic insulin release levels (right) of human islets as described in b. Data represent mean ± SD; n = 3 technical replicates; ***, P < 0.001; one-way ANOVA. (e) Schematic diagram of pancreatic ductal infusion of AAV and experimental design for f–j. (f) Representative image of pancreas and islets underwent pancreatic ductal infusion of AAV-GFP. Scale bar, 500 μm. (g) Immunoblots of total cell lysates from mouse islets from BKS-db/db mice 5 wk following AAV pancreatic ductal infusion. (h) Random blood glucose levels of BKS-db/db mice injected with saline or AAV-CTCF via pancreatic ductal infusion. Data represent mean ± SEM (n = 6 mice per group). ***, P < 0.001; two-way ANOVA. (i) Perifusion analyses of dynamic glucose-stimulated insulin secretion (left) and biphasic insulin release levels (right) of islets isolated from BKS-db/db mice as described in g. Each islet sample was pooled from at least three animals. Data represent mean ± SD; n = 3 technical replicates; ***, P < 0.001; two-tailed unpaired Student’s t test. (j) Glucose-stimulated Ca2+ influx in islets isolated from db/db mice as described in g. Each islet sample was pooled from at least three animals. Data represent mean ± SEM (n = 53–60 islets per group); ***, P < 0.001; two-way ANOVA. All panels report data verified in at least two independent experiments. Source data are available for this figure: SourceData F6.

AAV-mediated restore of CTCF expression ameliorates β cell dysfunction ex vivo and in vivo. (a) Schematic representation of experimental procedures for b–d. (b) Immunoblots of total cell lysates from mouse or human islets transduced with AAVs expressing CTCF or GFP for 5 d, followed by treatment with TNFα (50 ng/ml) and PA (0.5 mM) for an additional 24 h. (c) Perifusion analyses of dynamic glucose-stimulated insulin secretion (left) and biphasic insulin release levels (right) of mouse islets as described in b. Each islet sample was pooled from at least three animals. Data represent mean ± SD; n = 3 technical replicates; ***, P < 0.001; two-tailed unpaired Student’s t test. (d) Perifusion analyses of dynamic glucose-stimulated insulin secretion (left) and biphasic insulin release levels (right) of human islets as described in b. Data represent mean ± SD; n = 3 technical replicates; ***, P < 0.001; one-way ANOVA. (e) Schematic diagram of pancreatic ductal infusion of AAV and experimental design for f–j. (f) Representative image of pancreas and islets underwent pancreatic ductal infusion of AAV-GFP. Scale bar, 500 μm. (g) Immunoblots of total cell lysates from mouse islets from BKS-db/db mice 5 wk following AAV pancreatic ductal infusion. (h) Random blood glucose levels of BKS-db/db mice injected with saline or AAV-CTCF via pancreatic ductal infusion. Data represent mean ± SEM (n = 6 mice per group). ***, P < 0.001; two-way ANOVA. (i) Perifusion analyses of dynamic glucose-stimulated insulin secretion (left) and biphasic insulin release levels (right) of islets isolated from BKS-db/db mice as described in g. Each islet sample was pooled from at least three animals. Data represent mean ± SD; n = 3 technical replicates; ***, P < 0.001; two-tailed unpaired Student’s t test. (j) Glucose-stimulated Ca2+ influx in islets isolated from db/db mice as described in g. Each islet sample was pooled from at least three animals. Data represent mean ± SEM (n = 53–60 islets per group); ***, P < 0.001; two-way ANOVA. All panels report data verified in at least two independent experiments. Source data are available for this figure: SourceData F6.

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