Journal: Bioactive Materials

Article Title: Extracellular vesicles derived from salivary gland stem cells cultured on microwell scaffolds loaded with WNT3A promote the recovery of salivary gland function damaged by radiation via the YWHAZ-PI3K-AKT pathway

doi: 10.1016/j.bioactmat.2025.06.024

Figure Lengend Snippet: Functional validation of YWHAZ as a key effector in 3D WNT -EV-mediated restoration of irradiated mouse SGOs. (A) Representative bright-field, H&E, and PAS images of SGOs under five conditions: untreated (Control), irradiated + PBS (PBS), irradiated + 3D WNT -EVs derived from sgEpSCs transfected with YWHAZ siRNA (YWHAZ siRNA-EVs), irradiated + 3D WNT -EVs derived from sgEpSCs transfected with scrambled siRNA (scrambled siRNA-EVs), and irradiated + 3D WNT -EVs (3D WNT -EVs). SGOs treated with PBS or YWHAZ siRNA-EVs exhibited smaller size and cystic morphology, while those treated with scrambled siRNA-EVs or 3D WNT -EVs showed regenerative end-bud structures. H&E staining showed keratin pearl-like structures in the PBS group, while PAS staining revealed mucin recovery in all EV-treated groups, with greater intensity in scrambled and 3D WNT -EV conditions. Scale bars, 100 μm. (B) Representative immunofluorescence images of phosphorylated AKT (p-AKT) in SGOs under each treatment condition. SGOs treated with 3D WNT -EVs or scrambled siRNA-EVs showed elevated p-AKT levels compared to the PBS and YWHAZ siRNA-EVs groups. DAPI was used for nuclear counterstaining. Scale bars, 100 μm. (C) Cell viability, size of mouse SGOs and mucin area in EV-treated groups compared with the control and PBS groups. (D) Quantification of p-AKT + in SGOs shown in (B). Individual data points represent biological or technical replicates, per group ranging from 3 to 9. Error bars indicate SD. Statistical significance was determined using one-way ANOVA followed by Tukey's post hoc test. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: RNA interference (RNAi) was performed in human salivary gland–derived epithelial stem cells (sgEpSCs) using a Silencer® Pre-designed siRNA targeting human YWHAZ (ID: 107095, Ref: AM16708, Lot: ASO2NMXZ) and a non-targeting negative control siRNA (Ref: AM4611, Lot: ASO2NAC8), both purchased from Invitrogen (Thermo Fisher Scientific, USA).

Techniques: Functional Assay, Biomarker Discovery, Irradiation, Control, Derivative Assay, Transfection, Staining, Immunofluorescence

Journal: Scientific Reports

Article Title: Intestinal alkaline phosphatase is a receptor for cholesterol-lowering pentapeptide IIAEK and regulates cholesterol homeostasis in mice

doi: 10.1038/s41598-025-04722-w

Figure Lengend Snippet: IIAEK-human IAP complex interacts with cadherin-17. ( a ) SDS-PAGE analysis to identify the transmembrane target protein interacting with the human IAP- IIAEK (Ile-Ile-Ala-Glu-Lys) complex. ( b ) Identification of the transmembrane target protein interacting with the human IAP-IIAEK complex using nano LC–MS/MS and a Mascot search engine. ( c ) SDS-PAGE analysis to observe the interaction between the IIAEK-human IAP complex and cynomolgus cadherin-17. ( d ) SPR sensorgram of the interaction between cynomolgus cadherin-17 and immobilised human intestinal alkaline phosphatase (hIAP). ( e ) Effect of the addition of IIAEK on the SPR sensorgram of the interaction between cynomolgus cadherin-17 and immobilised hIAP. The original gels are presented in Supplementary Fig. 8.

Article Snippet: Subsequently, equal amounts of serum-free DMEM containing 240 nM CDH17 siRNA (Silencer ® Select Pre-designed siRNA Product, Ambion, 4392420, Thermo Fisher Scientific) or 240 nM Control siRNA (Silencncer ® Select Negative Control #1 siRNA, Ambion, 439084, Thermo Fisher Scientific) and serum-free DMEM containing siLentFectTM Lipid Reagent (BIO-RAD, 170–3361) were mixed and incubated at room temperature for 30 min to generate siRNA-siLentFect complexes.

Techniques: SDS Page, Liquid Chromatography with Mass Spectroscopy

Journal: Scientific Reports

Article Title: Intestinal alkaline phosphatase is a receptor for cholesterol-lowering pentapeptide IIAEK and regulates cholesterol homeostasis in mice

doi: 10.1038/s41598-025-04722-w

Figure Lengend Snippet: Effect of IIAEK on fluorescently labelled cholesterol uptake in Caco-2 cells with knockdown of human cadherin 17 ( CDH17 ). Caco-2 cells were cultured in 6-well plates for 14 days post-confluence. CDH 17 (20 nM) or negative control siRNA (20 nM) was introduced into the cells for 72 h. After 72 h of introduction, the cells were reintroduced with these siRNAs for 72 h. Subsequently, the cells were treated with serum-free medium containing 2 mM IIAEK (Ile-Ile-Ala-Glu-Lys) or the vehicle for 24 h. After removing the medium, the cells were incubated with a serum-free medium containing fluorescently labeled cholesterol (NBD-cholesterol) micelle for 1 h. After incubation, the cells were washed and cultivated to measure the fluorescent intensity of NBD-cholesterol. ( a ) CDH 17 mRNA levels. ( b ) Effect of IIAEK on NBD-cholesterol uptake in Caco-2 cells introduced with negative control siRNA. ( c ) Effect of IIAEK on NBD-cholesterol uptake in Caco-2 cells introduced with CDH17 siRNA. ( a – c ) n = 3/group. Data are presented as mean ± standard error of the mean (SEM). Statistical significance was calculated using an unpaired Student’s t -test. Asterisks indicate significant differences between the experimental groups (* p < 0.05).

Article Snippet: Subsequently, equal amounts of serum-free DMEM containing 240 nM CDH17 siRNA (Silencer ® Select Pre-designed siRNA Product, Ambion, 4392420, Thermo Fisher Scientific) or 240 nM Control siRNA (Silencncer ® Select Negative Control #1 siRNA, Ambion, 439084, Thermo Fisher Scientific) and serum-free DMEM containing siLentFectTM Lipid Reagent (BIO-RAD, 170–3361) were mixed and incubated at room temperature for 30 min to generate siRNA-siLentFect complexes.

Techniques: Knockdown, Cell Culture, Negative Control, Incubation, Labeling

Journal: Scientific Reports

Article Title: Intestinal alkaline phosphatase is a receptor for cholesterol-lowering pentapeptide IIAEK and regulates cholesterol homeostasis in mice

doi: 10.1038/s41598-025-04722-w

Figure Lengend Snippet: Hypothesis of how the specific interaction of IIAEK and IAP (IIAEK receptor) ameliorates cholesterol metabolism. The cholesterol-lowering pentapeptide IIAEK (Ile-Ile-Ala-Glu-Lys) specifically interacts with the GPI-anchored receptor intestinal alkaline phosphatase (IAP). Subsequently, the IIAEK-IAP dimer complex interacts with cadherin-17 to ameliorate cholesterol metabolism by suppressing cholesterol absorption, liver cholesterol, and serum cholesterol levels and promoting fecal cholesterol excretion.

Article Snippet: Subsequently, equal amounts of serum-free DMEM containing 240 nM CDH17 siRNA (Silencer ® Select Pre-designed siRNA Product, Ambion, 4392420, Thermo Fisher Scientific) or 240 nM Control siRNA (Silencncer ® Select Negative Control #1 siRNA, Ambion, 439084, Thermo Fisher Scientific) and serum-free DMEM containing siLentFectTM Lipid Reagent (BIO-RAD, 170–3361) were mixed and incubated at room temperature for 30 min to generate siRNA-siLentFect complexes.

Techniques: