human ift88  (Vector Biolabs)

Journal: bioRxiv

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

doi: 10.64898/2026.02.22.707280

Figure Lengend Snippet: ( 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–D ) Quantitative analysis of secretion traces. ( B ) Area under the curve (AUC) during liraglutide stimulation (minutes 30–50). ( C ) Total AUC of the entire perifusion (minutes 0-70). ( D ) AUC during KCl depolarization (minutes 60–70). βCKO islets secreted significantly less insulin in response to liraglutide and overall but showed no defect in KCl-induced secretion. Data are mean ± SEM. **p<0.01; ns, not significant, unpaired student’s t-test. ( E–J ) IFT88 knockdown impairs GLP-1-augmented secretion in human islets. Static glucose-stimulated insulin secretion (GSIS) assays from islets of male ( E–G ) and female ( H–J ) 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: Primary cilia regulate GLP-1 signaling in pancreatic β cells

doi: 10.64898/2026.02.22.707280

Figure Lengend Snippet: ( A ) Representative z-projected confocal images of human islets (28-year-old female donor) transduced with scrambled (Scr) or IFT88 -targeting shRNA adenovirus. GFP (green) marks transduced cells; ARL13B (red) labels primary cilia; DAPI (blue) stains nuclei. Scale bar = 20 µm. Cilia length was quantified from both conditions, showing significantly reduced mean length following IFT88 knockdown. n=132 cilia from 6 islets (Scr shRNA) and 467 cilia from 8 islets (IFT88 shRNA). ( B–C ) IFT88 knockdown efficiency was assessed across multiple donors to demonstrate reproducibility. ( B ) qPCR analysis of IFT88 mRNA levels in islets from two male donors (ages 27 and 44). ( C ) Immunoblot analysis of IFT88 protein levels in islets from three male donors (ages 36, 49 and 45), with corresponding quantifications. Bars represent scrambled control (gray) and IFT88 shRNA (pink). Data are mean ± SEM of duplicate or triplicate samples per donor. **p < 0.01, ****p < 0.0001, ns = not significant, by student’s unpaired t-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: Transduction, shRNA, Knockdown, Western Blot, Control

Journal: Frontiers in Molecular Biosciences

Article Title: Dendritic cell proliferation by primary cilium in atopic dermatitis

doi: 10.3389/fmolb.2023.1149828

Figure Lengend Snippet: IFT88 downregulation increases maturation marker expression with attenuating proliferation activity. (A) cDCs were stimulated with 1 μg/mL Df for 48 h. After immunostaining of acetylated tubulin, the percentage of primary ciliated cells was determined. n = 10 biological replicates (10 donors). The number of observed cells and primary cilium are as follows: no treatment; 8 cilia/1,074 cells. Df; 17 cilia/1,055 cells. The bar indicates the median value. **, p < 0.01 (Mann-Whitney U test). (B) cDCs were stimulated with 1

Article Snippet: After 5 days of differentiation, cells were electroporated with 20 nM siRNA targeting human IFT88 (Life Technologies), or siRNA Control (AM4611, Invitrogen) with the Neon Transfection System (Thermo Fisher Scientific).

Techniques: Marker, Expressing, Activity Assay, Immunostaining, MANN-WHITNEY, WST-1 Assay, Negative Control, Solvent, Comparison, Cell Culture, Control, Electroporation, Flow Cytometry, Derivative Assay, Western Blot

Journal: Acta Protozoologica

Article Title: The function of ciliopathy protein FOP on cilia and cortical microtubule cytoskeleton in Euplotes amieti

doi: 10.4467/16890027ap.23.005.18868

Figure Lengend Snippet: Figure 3. Protein loss and motility changes after RNAi. (A) Relative expression of basal body-associated protein BBS8. **P < 0.01 vs the blank group, n = 3. (B) Relative expression of cilia assembly-related protein IFT88. **P < 0.01 vs the blank group, n = 3. (C) Relative expression of basal body protein γ-tubulin. (D) The swimming tracks of Euplotes amieti during RNAi. The swimming speeds (n = 20). The FOPRNAi is the interference group, The Blank is the blank group (E) The swimming speeds of Euplotes amieti during RNAi (n = 20).

Article Snippet: Subsequently, the membranes were blocked in TBST buffer with 1 % skimmed milk for 2 hours at RT and respectively incubated with rabbit anti-human FOP (1:2000, Proteintech, China), rabbit anti-human β-tubulin (1:10000, Abcam, USA), rabbit anti-human BBS8 (1:500, Proteintech, China), rabbit anti-human IFT88 (1:1500, Proteintech, China), mouse anti-human γ-tubulin (1:10000, Sigma-Aldrich, USA), rabbit anti-human β-actin (1:10000, Proteintech, China) overnight at 4 °C, followed by peroxidase-conjugated sheep anti-rabbit/ mouse IgG secondary antibody (1:2000, Thermo Fisher Scientific, USA) for 2 hours at RT, respectively.

Techniques: Expressing