Fig. 2: The loss of CNOT3 in β cells impairs glucose tolerance and causes diabetes. | Communications Biology

Fig. 2: The loss of CNOT3 in β cells impairs glucose tolerance and causes diabetes.

From: Loss of β-cell identity and diabetic phenotype in mice caused by disruption of CNOT3-dependent mRNA deadenylation

Fig. 2

a Immunoblot analysis of CNOT3 in islet lysates from 8-week-old control and Cnot3βKO mice. This blot is a representative of three different blots. b Immunofluorescence of CNOT3 (green) in islets from 8-week-old control and Cnot3βKO mice. Nuclei were stained with DAPI (blue). A scale bar represents 50 µm. c Poly(A) tail lengths of global mRNAs by bulk poly (A) assay in control and Cnot3βKO islets (n = 1, each sample was pooled from four mice). d Densitograms of poly(A) tail lengths shown in (c). Intraperitoneal glucose tolerance tests in: e 4- and f 8-week-old control and Cnot3βKO mice (n = 6–9). g Fasting blood glucose in 12-week-old control (n = 5) and Cnot3βKO mice (n = 4). h Body weight comparisons between control and Cnot3βKO mice at 4, 8 and 12 weeks of age (n = 6). i Blood glucose in 8-week-old control (n = 6) and Cnot3βKO (n = 13) mice fed with ND. j Serum insulin in 8-week-old control and Cnot3βKO mice after 16 h fasting (0 min) and 15 min after glucose injection (2 g/kg body weight) (n = 4). k Insulin released by islets from 8-week-old control and Cnot3βKO mice in response to low 3 mM glucose (3G) and high 17 mM glucose (17G) stimulation for 1 h. This experiment is a representative of three independent experiments using three different biological replicates (n = 3). Data are presented as mean ± SEM; *P < 0.05; **P < 0.01; ***P < 0.001, two-tailed Student’s t test. l Cytosolic Ca2+ ([Ca2+] cyt) responses in control (n = 20) and Cnot3βKO (n = 18) islets from three 8-week-old mice from each genotype in response to low (3G), high (17G) glucose or 20 mM KCl. m AUC of glucose-evoked Ca2+ traces. AUC was calculated using different baseline values for each group (0.95 for control and 1.03 for Cnot3βKO). n AUC of depolarizing stimulus, KCl, response. AUC was calculated using different baseline values for each group (1.15 for control and 0.95 for Cnot3βKO). Data are presented as mean ± SEM; *P < 0.05; **P < 0.01; ***P < 0.001, Mann–Whitney test.

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