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: A neuropsychiatric disease-associated mutation in LRRC8B disrupts cellular Ca²⁺ signaling, mitochondrial function, and bioenergetics

doi: 10.64898/2026.04.16.718892

Figure Lengend Snippet: Volcano plot showing LC–MS/MS–based proteomic analysis of GFP pull-down samples from HEK293T cells expressing GFP-tagged wild-type (WT) or mutant LRRC8B. The x-axis represents log₂ fold change (mutant vs WT), and the y-axis shows −log₁₀ adjusted p-value. Proteins enriched in WT samples are shown on the left, while those enriched in mutant samples are shown on the right. Selected significantly enriched proteins are labeled. VDAC1 is identified as a WT-enriched interactor.

Article Snippet: The plasmid encoding GFP-tagged human LRRC8B (HG24935-ACG; Sino Biological Inc, USA).

Techniques: Liquid Chromatography with Mass Spectroscopy, Expressing, Mutagenesis, Labeling

Journal: bioRxiv

Article Title: A neuropsychiatric disease-associated mutation in LRRC8B disrupts cellular Ca²⁺ signaling, mitochondrial function, and bioenergetics

doi: 10.64898/2026.04.16.718892

Figure Lengend Snippet: Representative immunoblot images showing VDAC protein levels in total cell lysates and isolated mitochondrial fractions from cells transfected with GFP-tagged LRRC8B WT (W) or mutant Y380S (M) constructs. Actin (∼42 kDa) is included as a loading control for total lysates. Red boxes indicate the regions that were cropped and presented in the main figures. Molecular weight markers are shown where applicable.

Article Snippet: The plasmid encoding GFP-tagged human LRRC8B (HG24935-ACG; Sino Biological Inc, USA).

Techniques: Western Blot, Isolation, Transfection, Mutagenesis, Construct, Control, Molecular Weight

Journal: bioRxiv

Article Title: A neuropsychiatric disease-associated mutation in LRRC8B disrupts cellular Ca²⁺ signaling, mitochondrial function, and bioenergetics

doi: 10.64898/2026.04.16.718892

Figure Lengend Snippet: (A) Representative immunoblot images showing protein levels of GFP-tagged LRRC8B (WT and mutant) and VDAC in input lysates used for pull-down assays. (B–C) Immunoblot analysis of four independent immunoprecipitation (IP) experiments. Bands at ∼120 kDa confirm successful pull-down of GFP-tagged LRRC8B (WT and mutant) using an anti-GFP antibody. VDAC (∼32 kDa) bands indicate co-precipitation of VDAC with LRRC8B WT and mutant proteins. VDAC levels were normalized to the corresponding LRRC8B (WT or mutant) levels for quantification. Red boxes highlight the regions that were cropped and presented in the main figures. Molecular weight markers are shown where applicable.

Article Snippet: The plasmid encoding GFP-tagged human LRRC8B (HG24935-ACG; Sino Biological Inc, USA).

Techniques: Western Blot, Mutagenesis, Immunoprecipitation, Molecular Weight

Journal: bioRxiv

Article Title: Nup358 Sustains Intestinal Epithelial Homeostasis by Preventing Dvl1 Condensate Formation to Restrain Wnt Signaling

doi: 10.64898/2026.03.25.714063

Figure Lengend Snippet: a. Levels of the 7TGP fluorescent Wnt reporter in HEK293T cells transfected with control siRNA, Dvl1-specific siRNA, Nup358-specific siRNA, or a combination of Nup358-specific siRNA and Dvl1-specific siRNA and treated with either Wnt3a or vehicle. b. Western blot analysis and relative quantification of protein levels of Dvl1 and Axin1 in HEK293T cells transfected with either control or Nup358-specific siRNA. α-Tubulin and GAPDH were used as loading controls. Data are expressed as mean ± SD. **p ≤ 0.01. c. Immunofluorescence staining for Dvl1 and Nup358 in HEK293T cells transfected with either control or Nup358-specific siRNA. d. Immunofluorescence staining for Dvl1 and Nup358 in HEK293T cells transfected with control or Nup358-specific siRNA and treated with vehicle or 1,6-hexanediol 5% for 2 minutes. e. Fluorescence recovery after photobleaching (FRAP) and fusion analysis of Dvl1 condensates in HEK293T cells transfected with Nup358-specific siRNA and EGFP-Dvl1.

Article Snippet: Plasmids used in this study include β-catenin-EGFP (Addgene, #71367), EGFP-Dvl1 (Addgene, #194586) and plasmids HA-Nup358 1–2683 , HA-Nup358 1–2148 , HA-Nup358 1–1810 , HA-Nup358 1–1133 from Wälde et al 13 .

Techniques: Transfection, Control, Western Blot, Quantitative Proteomics, Immunofluorescence, Staining, Fluorescence

Journal: bioRxiv

Article Title: Nup358 Sustains Intestinal Epithelial Homeostasis by Preventing Dvl1 Condensate Formation to Restrain Wnt Signaling

doi: 10.64898/2026.03.25.714063

Figure Lengend Snippet: a. Schematic representing different functional domains of Nup358. b. Fluorescent Wnt reporter activity in HEK293T cells transfected with control siRNA, Nup358-specific siRNA, Ubc9-specific siRNA, or treated with the SUMOylation inhibitor 2-D08. c. Schematic representing different HA-tagged truncated forms of Nup358 that were transiently expressed in HEK293T cells. d. Fluorescent Wnt reporter activity in HEK293T cells transfected with either control or Nup358-specific siRNA alone or in combination with different HA-tagged truncated forms of Nup358 were analyzed by confocal microscopy (top) and quantified (bottom). Data are expressed as mean ± SD. ****p ≤ 0.0001 e. Western blot analysis of co-immunoprecipitation of HA-Nup358 and EGFP-Dvl1 transiently expressed in HEK293T cells. GAPDH was used as loading control. f. Immunofluorescence of HEK293T cells transfected with HA-Tagged Nup358 1–1133 and EGFP-Dvl1and stained with an anti-HA antibody. g. Immunofluorescence staining for endogenous Dvl1 and Nup358 in HEK293T cells transfected with either scramble control or Nup358-specific siRNA alone or in combination with HA-Tagged Nup358 1–1133 fragment were analyzed by confocal microscopy (top) and quantified (bottom). Data are expressed as mean ± SD. **p ≤ 0.01.

Article Snippet: Plasmids used in this study include β-catenin-EGFP (Addgene, #71367), EGFP-Dvl1 (Addgene, #194586) and plasmids HA-Nup358 1–2683 , HA-Nup358 1–2148 , HA-Nup358 1–1810 , HA-Nup358 1–1133 from Wälde et al 13 .

Techniques: Functional Assay, Activity Assay, Transfection, Control, Confocal Microscopy, Western Blot, Immunoprecipitation, Immunofluorescence, Staining