Journal: International journal of molecular sciences

Article Title: Modulation of Unfolded Protein Response Restores Survival and Function of β-Cells Exposed to the Endocrine Disruptor Bisphenol A.

doi: 10.3390/ijms24032023

Figure Lengend Snippet: Figure 3. Insulin processing is affected by BPA and Tm in MIN6 cells and isolated islets. (A) Time course of BPA-induced Ins2 gene expression at different time points (2, 6, 12, 24, 48 h) in MIN6 cells. RT-qPCR analysis of the Ins2 gene in MIN6 cells (B) and isolated islets (C) after 24 h of treatment with different concentrations of BPA and 5 µg/mL Tm. Fold change values were calculated by normalization to Gapdh and then to the NT values. (D) Representative fluorescence microscopy images with MIN6 cells after 24 h of treatment with 100, 250 µM BPA and 5 µg/mL Tm, marked for proinsulin (red) and insulin (green). The nucleus was stained with DAPI. (E) Graphic representation of the mean intensity of the red fluorescent channel (proinsulin) and (F) the mean intensity of the green fluorescent channel (insulin) per cell (n > 200 cells) in MIN6 cells after 24 h of treatment with increasing concentrations of BPA (as shown) and 5 µg/mL Tm. The mean intensity was determined with QuPath. Graphs display the median and the 25th and 75th percentiles. * p < 0.05, **** p < 0.0001, ns—not significant, based on one way ANOVA.

Article Snippet: The following antibodies were used for immunofluorescence: mouse monoclonal antihuman/mouse proinsulin biotinylated antibody (1:200; R&D Systems (Minneapolis, MN, USA), #BAM13361), mouse monoclonal anti-calnexin antibody (1:100; Novus Biologicals (Englewood, CO, USA), #NB300-518), rabbit polyclonal anti-GM130 antibody (1:200; Novus Biologicals, #NBP2-53420), guinea pig polyclonal anti-insulin antibody (1:50; GeneTex (Zeeland, MI, USA), #GTX27842).

Techniques: Isolation, Gene Expression, Quantitative RT-PCR, Microscopy, Staining

Journal: Molecular Metabolism

Article Title: Beta-cell-specific C3 deficiency exacerbates metabolic dysregulation and insulin resistance in obesity

doi: 10.1016/j.molmet.2025.102302

Figure Lengend Snippet: A) Pancreas sections from treated mice were stained for glucagon (green) and insulin (red) to visualize islets and assess their size. Blue: DAPI. Scale bars, 50 μm. B) Quantification of total islet size in sections from 22 islets from pancreas sections of 7 LFD beta-C3-KO mice, 33 islets from pancreas sections of 5 HFD beta-C3-KO mice, and 35 islets from pancreas sections of 6 HFD C3-flox mice. C) Fraction of total islet staining for insulin, from the same islets analysed in (B). D) Examples of staining of individual HFD islets for glucagon (green) and pro-insulin (red). E) Quantification of pro-insulin staining in islets from each group of mice. F) Quantification of pro-insulin in serum of mice at given time points, measured by pro-insulin specific ELISA. G) Transmission electron micrographs of pancreatic islet cells isolated from beta-C3-KO or C3-flox mice. Beta-C3-KO mouse beta-cells contain accumulated amounts of swollen ER or similar vesicular organelles (black arrows), while normal ER is apparent in C3-flox mouse islet cells (white arrows). See also . Increased amounts of autophagic material were also observed in beta-C3-KO cells (asterisk). H) In vitro insulin secretion from isolated islets from C3-flox or beta-C3-KO mice HFD-fed mice, at 15 or 60 min. I) Average insulin content values per islet from secretion experiment in panel (H) (average values shown for each of 6 mice: from 6 wells, 5 islets per well). J) Serum C3 levels in C3-flox or beta-C3-KO mice at the endpoint of HFD, as measured by ELISA. Statistics in E, one-way ANOVA, in F/H, two-way ANOVA, in I, Student’s T-test, and in B/C, by mixed model statistical analysis (see methods).

Article Snippet: For pro-insulin staining, AlexaFluor647-labeled anti-pro-insulin (R&D Systems, #IC13361R, 1:100) was incubated overnight at 4 °C, in combination with glucagon staining, before washing and mounting as described above.

Techniques: Staining, Enzyme-linked Immunosorbent Assay, Transmission Assay, Isolation, In Vitro

Journal: iScience

Article Title: β-chitosan attenuates hepatic macrophage-driven inflammation and reverses aging-related cognitive impairment

doi: 10.1016/j.isci.2024.110766

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-IL-6 , Abmart , Cat#TD6087.

Techniques: Plasmid Preparation, Recombinant, Lysis, Membrane, Staining, Enzyme-linked Immunosorbent Assay, Bicinchoninic Acid Protein Assay, Western Blot, Software, Expressing, Gene Expression