Journal: ACS Applied Nano Materials

Article Title: CdSe Quantum Dot-Based Delivery System for CRISPR-Cas9 Mediated Microglial Gene Modulation

doi: 10.1021/acsanm.4c04586

Figure Lengend Snippet: Figure 7. Stability and release of Cas9/Qdot conjugates. (a) Confocal microscopy images of BV2 cells treated with Cas9/Qdot conjugates for 6− 72 h. Nuclei were stained with DAPI. Scale bar = 10 μm. (b) Fluorescence intensity of the released Cas9 proteins per cell. (c) In vitro analysis of Cas9 releasing using 300 kDa cutoff filtering. SDS-PAGE analysis (d) and quantification (e) of conjugates containing maleimide-biotin linker or GSH. Data are presented as mean ± standard deviation; N = 5 independent ROIs (b,e). **p < 0.01, ***p < 0.001, ****p < 0.0001, # not significant vs at 6 h (b) and 0 h (e); assessed using one-way ANOVA with Dunnett’s multiple comparisons test. Qdot, quantum dot; DAPI, 4′,6- diamidino-2-phenylindole; h, hours; ROI, regions of interest; GSH, glutathione.

Article Snippet: For stability and release evaluation, the conjugates were incubated at 37 °C with 1 mM reduced glutathione (GSH; Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Techniques: Confocal Microscopy, Staining, Fluorescence, In Vitro, SDS Page, Standard Deviation

Journal: ACS Applied Nano Materials

Article Title: CdSe Quantum Dot-Based Delivery System for CRISPR-Cas9 Mediated Microglial Gene Modulation

doi: 10.1021/acsanm.4c04586

Figure Lengend Snippet: Figure 8. Intracellular fate of Cas9/Qdot conjugates in microglia. The conjugates are released by free thiols (ex. GSH) in the cytoplasm at 24 h after selective internalization by CME. Although disassembled Cas9 proteins are prone to degradation by proteases, they can also pass through the nuclear pore and edit DNA targets (ex. Rosa26, HPRT1). In addition, the released Qdots can exert anti-inflammatory effects via radical scavenging. Qdot, quantum dot; h, hours; GSH, glutathione; CME, clathrin-mediated endocytosis.

Article Snippet: For stability and release evaluation, the conjugates were incubated at 37 °C with 1 mM reduced glutathione (GSH; Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Techniques:

Journal: Molecules

Article Title: Biotransformation of Timosaponin BII into Seven Characteristic Metabolites by the Gut Microbiota

doi: 10.3390/molecules26133861

Figure Lengend Snippet: The structure and mass spectra of timosaponin BII. ( A ) The structural formula of timosaponin BII and the Glipizide (internal standard (IS)). ( B ) Extracted ion chromatogram (EIC) spectra of timosaponin BII and the internal standard (Glipizide). ( C ) The high-resolution mass spectra of timosaponin BII acquired by LC/MS-Q-TOF.

Article Snippet: Timosaponin BII (CAS, 136656-07-0; Cat Number, IT0640), timosaponin AIII (CAS, 41059-79-4; Cat Number, IT0630), Sarsasapogenin (CAS, 126-19-2; Cat Number, SS8160), and Glipizide (CAS, 29094-61-9; Cat Number, SG8680) were purchased from Solarbio Life Sciences Co., Ltd. (Beijing, China).

Techniques: Liquid Chromatography with Mass Spectroscopy

Journal: Molecules

Article Title: Biotransformation of Timosaponin BII into Seven Characteristic Metabolites by the Gut Microbiota

doi: 10.3390/molecules26133861

Figure Lengend Snippet: The level of possible timosaponin BII metabolites changed during incubation with rat intestinal bacteria after 0 min, 15 min, 30 min, 60 min, 90 min, and 120 min. ( A ) Metabolic curve of timosaponin AIII. ( B ) Metabolic curve of timosaponin AI. ( C ) Metabolic curve of sarsasapogenin. ( D ) The temporal characteristics of potential timosaponin BII metabolites by gut microbiota. ( E ) Extracted ion chromatogram (EIC) spectra of timosaponin AIII, timosaponin AI, sarsasapogenin and the internal standard (Glipizide). ( F ) Metabolic pathway of timosaponin BII and possible structures of metabolites.

Article Snippet: Timosaponin BII (CAS, 136656-07-0; Cat Number, IT0640), timosaponin AIII (CAS, 41059-79-4; Cat Number, IT0630), Sarsasapogenin (CAS, 126-19-2; Cat Number, SS8160), and Glipizide (CAS, 29094-61-9; Cat Number, SG8680) were purchased from Solarbio Life Sciences Co., Ltd. (Beijing, China).

Techniques: Incubation