strains mic ml mic ml sy neo teo neo teo l innocua  (ATCC)

Journal: Islets

Article Title: Senp2 expression was induced by chronic glucose stimulation in INS1 cells, and it was required for the associated induction of Ccnd1 and Mafa

doi: 10.1080/19382014.2016.1235677

Figure Lengend Snippet: Expression of Senp2 compared to Senp1 . (A) Human islets were isolated and stabilized in the RPMI containing 10% FBS overnight. The mRNA expression of SENP1 and SENP2 was measured by quantitative RT-PCR and presented as ratios to ACTIN (n = 5). (B) IHC staining for SENP2 (brown) was performed on human pancreas harvested from a non-DM and a type 2 DM patients. Islets are indicated by dotted lines (upper panels) and presented on higher magnification (lower panels). (C) INS1 cells were cultured in RPMI containing 10% FBS, and either 0.25 mM palmitic acid or 25 mM high glucose was added for 72 h. The expression of Senp1 and Senp2 mRNA was measured by quantitative RT-PCR, and was presented as ratios to Actin expression (n = 5). Student's t-test was used for the comparisons between Senp1 and Senp2 . DM, diabetes mellitus; NS, no significant difference; PA, palmitic acid.

Article Snippet: When INS1 cells reached 70∼80% confluence, they were transfected with si Senp2 (Santa Cruz Biotechnology, sc-72204) or a negative control small interfering RNA (siNS) (Bioneer, SN-1003) at 200 nM using Lipofectamine RNAiMAX (Thermo Fisher Scientific, #56532).

Techniques: Expressing, Isolation, Quantitative RT-PCR, Immunohistochemistry, Cell Culture

Journal: Islets

Article Title: Senp2 expression was induced by chronic glucose stimulation in INS1 cells, and it was required for the associated induction of Ccnd1 and Mafa

doi: 10.1080/19382014.2016.1235677

Figure Lengend Snippet: Expression of Senp2 in the islets of hyperglycemic mice. (A) Male C57BL/6 wild-type mice were fed with normal chow or HFD for 8 weeks, with monitoring of blood glucose and body weight every week. Male ob/ob , and db/db mice at the same age were monitored, too. Numbers of animals were 2 in the each group. (B-D) H&E stain sections and IHC staining (brown color) for Insulin and Senp2 of pancreas from each group. H&E, hematoxylin and eosin; HFD, high fat diet; IHC, immunohistochemical.

Article Snippet: When INS1 cells reached 70∼80% confluence, they were transfected with si Senp2 (Santa Cruz Biotechnology, sc-72204) or a negative control small interfering RNA (siNS) (Bioneer, SN-1003) at 200 nM using Lipofectamine RNAiMAX (Thermo Fisher Scientific, #56532).

Techniques: Expressing, Staining, Immunohistochemistry, Immunohistochemical staining

Journal: Islets

Article Title: Senp2 expression was induced by chronic glucose stimulation in INS1 cells, and it was required for the associated induction of Ccnd1 and Mafa

doi: 10.1080/19382014.2016.1235677

Figure Lengend Snippet: Expression of Senp2 and cell number according to the duration of high glucose in INS1 cells. INS1 cells were incubated in the control media (RPMI containing 10% FBS and 11 mM glucose) and high-glucose (25 mM) media for the indicated time, and the sequential changes were studied. (A) Senp2 mRNA was measured by quantitative RT-PCR, and was presented as ratios to Actin expression (n = 3). (B) Senp2 protein levels were measured by the western blotting, and presented as ratios to Actin expression (n = 4). (C) Cell number was presented as a fold of seeded cell number (n = 3). (D) Expression of Ccnd1 transcript was measured by quantitative RT-PCR (n = 3). (E) S6K1 phosphorylation was detected by the western blotting and presented as ratios to total S6K1 expression (n = 3). (F) Caspase 3 cleavage was examined by the western blotting and presented as ratios to intact Caspase 3 (n = 4). Open bars, control media with 11 mM of glucose; solid bars, high-glucose (25 mM) media. Representative protein gel blots are below the graphs for B, E & F. In A, C, D and E, 2-way repeated measures ANOVA and Bonferroni posttests were performed. In B and F, paired t-test was performed. *, p < 0.05; **, p < 0.01; NS, no significant difference.

Article Snippet: When INS1 cells reached 70∼80% confluence, they were transfected with si Senp2 (Santa Cruz Biotechnology, sc-72204) or a negative control small interfering RNA (siNS) (Bioneer, SN-1003) at 200 nM using Lipofectamine RNAiMAX (Thermo Fisher Scientific, #56532).

Techniques: Expressing, Incubation, Control, Quantitative RT-PCR, Western Blot, Phospho-proteomics

Journal: Islets

Article Title: Senp2 expression was induced by chronic glucose stimulation in INS1 cells, and it was required for the associated induction of Ccnd1 and Mafa

doi: 10.1080/19382014.2016.1235677

Figure Lengend Snippet: Suppression of Senp2 during high glucose and associated changes in Ccnd1, Mafa and cell number. INS1 cells at 80% confluence were transfected with siNS and si Senp2 . After overnight incubation, the media was changed with and without high glucose (25 mM) for 48 and 72 h (n = 4). The ratios of each transcript to Actin measured by quantitative RT-PCR were presented as relative values compared to the control at 48 h (A) and at 72 h (B). (C) Cell number was compared to the control at 48 h and at 72 h. One-way ANOVA and Tukey's Multiple Comparison Test were used. *, p < 0.05; **, p < 0.01; NS, no significant difference.

Article Snippet: When INS1 cells reached 70∼80% confluence, they were transfected with si Senp2 (Santa Cruz Biotechnology, sc-72204) or a negative control small interfering RNA (siNS) (Bioneer, SN-1003) at 200 nM using Lipofectamine RNAiMAX (Thermo Fisher Scientific, #56532).

Techniques: Transfection, Incubation, Quantitative RT-PCR, Control, Comparison

Journal: Islets

Article Title: Senp2 expression was induced by chronic glucose stimulation in INS1 cells, and it was required for the associated induction of Ccnd1 and Mafa

doi: 10.1080/19382014.2016.1235677

Figure Lengend Snippet: Overexpression of h SENP2 and associated changes in Ccnd1, Insulin and cell number. INS1 cells at 80% confluence were infected with Ad-h SENP2 and Ad-GFP. After overnight incubation, the media was changed with and without high glucose (25 mM) for 48 h (n = 8∼9). (A) A representative picture of western blotting. (B) The ratios of each transcript to Actin were presented as relative values compared to the control. h SENP2 was detected only in the cells infected by Ad-h SENP2 . (C) Cell number was compared to the control. Paired t-test and Wilcoxon matched-pairs signed rank test were used between the indications. *, p < 0.05; NS, no significant difference.

Article Snippet: When INS1 cells reached 70∼80% confluence, they were transfected with si Senp2 (Santa Cruz Biotechnology, sc-72204) or a negative control small interfering RNA (siNS) (Bioneer, SN-1003) at 200 nM using Lipofectamine RNAiMAX (Thermo Fisher Scientific, #56532).

Techniques: Over Expression, Infection, Incubation, Western Blot, Control