Journal: Diabetologia

Article Title: Saturated fatty acids entrap PDX1 in stress granules and impede islet beta cell function.

doi: 10.1007/s00125-021-05389-4

Figure Lengend Snippet: Fig. 2 SFAs sequester PDX1 in SGs. (a) Distribution of the identified proteins according to molecular function. (b) INS1 cells were treated with PA (0.4 mmol/l) for 16 h and then stained with anti-TIA1 antibody (green), anti- PDX1 antibody (red) and DAPI (blue). Short arrows indicate co- localisation. Scale bar, 5 μm. On the right, line scans are shown of the images of a cell co-stained for G3BP1 and PDX1 at the position depicted by the long arrow. (c, d) PDX1 levels in the SG-enriched fraction separated from INS1 cells (c) or human islets (d), as well as ataxin2 and G3BP1 in (c) and TIA1 in both. (e, f) Co- immunoprecipitation of SG markers, including ataxin2, G3BP1 and TIA1 with PDX1 from INS1 cells treated with PA for 16 h (e), or from mouse pancreatic tissues after HFD for 10 weeks (f). Data are presented as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001. (c–e) n = 3; (f) n = 6. Ctrl, control; IP, immunoprecipitate; NC, normal chow; P18000, pellet from 18,000 × g; ROI, region of interest; S, supernatant after 18,000 × g

Article Snippet: Materials Anti-PDX1 (catalogue no. #5679), anti-phosphoPI3 kinase p85 (Tyr458)/p55 (Tyr199) (catalogue no. #17366) antibodies and DAPI (catalogue no. #4083) were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Staining, Immunoprecipitation, Control

Journal: Diabetologia

Article Title: Saturated fatty acids entrap PDX1 in stress granules and impede islet beta cell function.

doi: 10.1007/s00125-021-05389-4

Figure Lengend Snippet: Fig. 4 Suppression of SG formation by G3bp1 or Tia1 knockdown restored pancreatic function. (a, b) INS1 cells were transfected with Tia1 siRNA (a) or G3bp1 siRNA (b) for 72 h and WCL were subjected to immunoblotting with the indicated antibodies. (c) INS1 cells were transfected with Tia1 siRNA for 72 h, followed by PA (0.4 mmol/l) treatment for 16 h, cells were fixed and stained for G3BP1 (red) and DAPI (blue). Quantitative analysis of the distribution of SGs in INS1 cells is shown on the right. Scale bar, 10 μm. (d, e) INS1 cells were transfected with the indicated siRNA for 72 h, followed by PA treatment

Article Snippet: Materials Anti-PDX1 (catalogue no. #5679), anti-phosphoPI3 kinase p85 (Tyr458)/p55 (Tyr199) (catalogue no. #17366) antibodies and DAPI (catalogue no. #4083) were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Knockdown, Transfection, Western Blot, Staining