anti phospho mtor  (Proteintech)

Journal: Materials Today Bio

Article Title: Renal-targeted tFNA-TPP nanoagonist treats acute kidney injury by amplifying mitophagy

doi: 10.1016/j.mtbio.2026.102926

Figure Lengend Snippet: Design of tFNA-TPP and its role in targeted mitochondrial clearance in the kidney. (A) Synthesis of alkynyl-modified primer (AP), which is conjugated with 3-azidopropyl triphenyl phosphine (TPP) via Cu(I)-catalyzed Huisgen cycloaddition to form AP-TPP. (B) Preparation of a Cy5-labeled primer (Cy5-P) and four single-stranded DNAs (S1, S2, S3, S4) such that Cy5-P is complementary to the 5′ end of S4, while AP-TPP is complementary to the 5′ ends of S1, S2, and S3. (C) Self-assembly of the four DNA strands (S1, S2, S3, S4) leads to the formation of a tetrahedral framework nucleic acid (tFNA), which is further functionalized with AP-TPP and Cy5-P to yield tFNA-TPP. (D) tFNA-TPP is applied for the targeted delivery to damaged mitochondria in renal cells. This approach promotes the clearance of damaged mitochondria by modulating autophagy-related proteins (upregulation of LC3-2, downregulation of P62 and COX4), thereby alleviating mitochondrial damage associated with acute kidney injury (AKI).

Article Snippet: Nonspecific binding sites were blocked by incubating the sections with goat serum for 1 h. After washing, the samples were incubated overnight at 4 °C with diluted primary antibodies against TOM20 (Proteintech, China) and LC3 (Proteintech, China), respectively.

Techniques: Modification, Labeling

Journal: Materials Today Bio

Article Title: Renal-targeted tFNA-TPP nanoagonist treats acute kidney injury by amplifying mitophagy

doi: 10.1016/j.mtbio.2026.102926

Figure Lengend Snippet: Assessment of tFNA-TPP's efficacy in ROS clearance and mitophagy induction. (A) Fluorescence microscopy images showing ROS levels in cells treated for 24 and 48 h: (I) Saline, (II) H 2 O 2 , (III) H 2 O 2 + tFNA, (IV) H 2 O 2 + AP-TPP, and (V) H 2 O 2 + tFNA-TPP. A significant reduction in ROS is observed in the tFNA-TPP group over time. (B) Schematic depicting the dual role of tFNA-TPP in reactive oxygen species (ROS) clearance and facilitation of mitophagy. (C) Confocal images illustrating mitophagy in cells expressing GFP-LC3 and mRFP-LC3 treated under different conditions, with enhanced colocalization in the tFNA-TPP group. (D) Quantification of autophagosome-to-autolysosome conversion ratio, showing highest efficacy in group V (tFNA-TPP), n = 3. (E) Ratio of experimental to control group metrics validating the enhanced performance of tFNA-TPP, n = 3. (F) Transmission electron microscopy (TEM) images revealing mitochondrial ultrastructure changes, with significant autophagic activity observed in group V. (G) Western blot analysis of autophagy markers, presenting decreased P62 and increased LC3-II conversion, and reduced COX4 levels in the tFNA-TPP treated group, indicating enhanced mitophagic activity.

Article Snippet: Nonspecific binding sites were blocked by incubating the sections with goat serum for 1 h. After washing, the samples were incubated overnight at 4 °C with diluted primary antibodies against TOM20 (Proteintech, China) and LC3 (Proteintech, China), respectively.

Techniques: Fluorescence, Microscopy, Saline, Expressing, Control, Transmission Assay, Electron Microscopy, Activity Assay, Western Blot

Journal: Materials Today Bio

Article Title: Renal-targeted tFNA-TPP nanoagonist treats acute kidney injury by amplifying mitophagy

doi: 10.1016/j.mtbio.2026.102926

Figure Lengend Snippet: Therapeutic evaluation of tFNA-TPP in a cisplatin-induced acute kidney injury (AKI) model. (A) Experimental schematic depicting cisplatin administration to induce AKI, followed by tFNA-TPP treatment and subsequent collection of kidneys and blood at 72 h. (B) Serum creatinine (CRE) levels indicate substantial renal function preservation in the tFNA-TPP group (V) compared to cisplatin alone (II). (C) Blood urea nitrogen (BUN) levels corroborate improved renal function with tFNA-TPP intervention. Relative ATP levels highlight improved cellular energy states in tFNA-TPP treated mice. (D) Malondialdehyde (MDA) levels, indicating oxidative stress, show a significant reduction with tFNA-TPP treatment. (E) Superoxide dismutase (SOD) activity analysis reveals enhanced antioxidant defense in the tFNA-TPP treated group. (F) Relative ATP levels highlight improved cellular energy states in tFNA-TPP treated mice. (G) Total glutathione (GSH) levels reflect an increased antioxidant capacity in the tFNA-TPP group. (H) Histological evaluation with hematoxylin and eosin (HE), periodic acid-Schiff (PAS), KIM-1 immunostaining, and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assays reveal significant reduction in injury and apoptosis, and restoration of renal architecture in the tFNA-TPP group. × 20. (I) Western blot analysis of HO-1, Nrf2, and cleaved caspase-3 expressions further confirms reduced oxidative stress and apoptosis in the tFNA-TPP treated group, supporting its protective effects against cisplatin-induced kidney damage. (J) Transmission electron microscopy (TEM) images depict renal ultrastructural improvement with reduced mitochondrial and cellular damage in the tFNA-TPP treated group. (K) Western blot analysis showing downregulation of P62 and COX4, and upregulation of LC3-II/LC3-I ratio, indicating enhanced autophagic response in tFNA-TPP treated kidneys.

Article Snippet: Nonspecific binding sites were blocked by incubating the sections with goat serum for 1 h. After washing, the samples were incubated overnight at 4 °C with diluted primary antibodies against TOM20 (Proteintech, China) and LC3 (Proteintech, China), respectively.

Techniques: Preserving, Activity Assay, Immunostaining, TUNEL Assay, Western Blot, Transmission Assay, Electron Microscopy

Journal: Materials Today Bio

Article Title: 3D-printed titanium scaffolds coated with a multifunctional photothermal-responsive hydrogel promote osteoporotic bone defect repair

doi: 10.1016/j.mtbio.2026.102879

Figure Lengend Snippet: Effects of hydrogel coatings on delaying senescence of BMSCs (A) Representative image of SA- β -gal staining. (B) Quantitative analysis of SA- β -gal staining intensity. (C) mRNA expression levels of p16 , p53 , Ccl2 , Tnf-α , IL-1β, and IL-6 in BMSCs. (D) Quantitative analysis of p21 fluorescence intensity. (E) Representative fluorescence images of p62 staining. (F) Quantitative analysis of p62 fluorescence intensity. (G) Representative fluorescence images of LC3B staining. (H) Quantitative analysis of LC3B fluorescence intensity. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001, n = 3.

Article Snippet: Primary antibodies against OPN (22952-1-AP), RUNX2 (20700-1-AP), CD86 (13395-1-AP), CD206 (18704–1-AP), p21 (10355-1-AP), p16 (10883-1-AP), LC3B (14600-1-AP), and CL594-phalloidin (PF00003) were obtained from Proteintech (Wuhan, China).

Techniques: Staining, Expressing, Fluorescence