human ift88  (Vector Biolabs)

Journal: Molecular Metabolism

Article Title: Primary cilia regulate GLP-1 signaling in pancreatic β cells

doi: 10.1016/j.molmet.2026.102357

Figure Lengend Snippet: Primary cilia are required for GLP-1-potentiated insulin secretion. ( A ) Dynamic insulin secretion from perifused mouse islets. Wild-type (WT, black) and β-cell-specific cilia knockout (βCKO, red) islets were sequentially exposed to 2 mM glucose (2G), 16 mM glucose (16G), 20 nM liraglutide (Lira), and 30 mM KCl as indicated. n = 6 replicates of 50 islets per genotype from 5 mice. ∗p < 0.05, two-way ANOVA with Sidak's multiple comparisons test. ( B ) Quantitative analysis of secretion traces: area under the curve (AUC) during initial glucose stimulation (minutes 10–30), liraglutide stimulation (minutes 30–50), KCl depolarization (minutes 60–70), and total AUC of the entire perifusion (minutes 0–70). βCKO islets secreted significantly less insulin in response to glucose and liraglutide but showed no defect in KCl-induced secretion. Data are mean ± SEM. ∗∗p < 0.01; ns, not significant, unpaired student's t-test. ( C-D ) IFT88 knockdown impairs GLP-1-augmented secretion in human islets. Static glucose-stimulated insulin secretion (GSIS) assays from islets of three male ( C ) and three female ( D ) donors. Islets transduced with control (Ctl, white) or IFT88 -targeting shRNA (IFT88 KD, red) were incubated at 1 mM glucose (1G), 11 mM glucose (11G), and 11G + 100 nM liraglutide (Lira). Donor ages are indicated. IFT88 KD significantly reduced liraglutide-potentiated insulin secretion. Data are mean ± SEM of triplicate samples per donor. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001; ns, not significant; one-way ANOVA with Tukey's multiple comparisons test.

Article Snippet: IFT88 knockdown in human islets: Healthy non-diabetic human islets were transduced with adenoviral vectors encoding either GFP-tagged shRNA targeting human IFT88 (Ad-GFP-h-IFT88-shRNA) or scrambled control (Ad-GFP-U6-scrmb-shRNA, #1122N; Vector Biolabs).

Techniques: Knock-Out, Knockdown, Transduction, Control, shRNA, Incubation

Journal: bioRxiv

Article Title: Phosphoproteomics identifies the DYRK1B protein kinase as a regulator of processing bodies

doi: 10.64898/2026.04.29.721300

Figure Lengend Snippet: (A) Time course induction of FLAG-DYRK1B in HD1B cells using Tetracycline (1µM) for up to 24h shows increased phosphorylation of DCP1A and 4E-T targets. Phosphorylation coincides with the expression of DYRK1B protein. (B) Expression of WT but not kinase dead (KD) DYRK1B, upon Tetracycline (1µM) treatment for 24h, phosphorylates DCP1A, 4E-T and PAT1B in HD1B cells. This phosphorylation is inhibited if cells are treated with the DYRK1B inhibitor AZ191 (1µM). (C) Phos-tag gel showing the shift of phosphorylated 4E-T, DCP1A, PAT1B, EDC3 proteins upon DYRK1B expression and the impaired phosphorylation upon treatment with AZ191 (1µM). (D) Expression of WT but not kinase dead (K140R and D239A mutants) EGFP-DYRK1B in HeLa Flp-In T-Rex cells increases phosphorylation of DCP1A and p62 (T269/S272). (E) Knockdown of DYRK1B in PANC-1 cells using siRNA reduces DCP1A and 4E-T phosphorylation.

Article Snippet: In a 12-well format, 0.5μg DNA was mixed with 75μl JetPrime buffer and 2μl reagent, incubated for 10 min at room temperature, added to cells for 6 h, and then replaced with fresh complete medium for a further 24–48 h. For knockdown experiments, GFP siRNA (Eurofins), a negative control siRNA (Thermo Fisher Scientific), ON-TARGETplus human DYRK1B siRNA (Dharmacon) and Silencer siDYRK1B (Thermo Fisher Scientific) were used at a final concentration of 30nM. siRNAs were transfected using Lipofectamine RNAiMAX according to the manufacturer’s instructions.

Techniques: Phospho-proteomics, Expressing, IF-cells, Knockdown