Journal: Cell reports

Article Title: CD14 is a decision-maker between Fas-mediated death and inflammation

doi: 10.1016/j.celrep.2024.114685

Figure Lengend Snippet:

Article Snippet: Caspase 3/7 inhibitor , Cayman Chemical Company , Cat#14464.

Techniques: Recombinant, Reverse Transcription, CyQUANT Assay, LDH Cytotoxicity Assay, In Situ, TUNEL Assay, Enzyme-linked Immunosorbent Assay, Plasmid Preparation, Software

Journal: Nature methods

Article Title: Mapping Enzyme Activity in Living Systems by Real-Time Mid-Infrared Photothermal Imaging of Nitrile Chameleons

doi: 10.1038/s41592-023-02137-x

Figure Lengend Snippet: (a) Schematic illustration of the principle of enzyme activity mapping by real-time MIP imaging of nitrile chameleons. (b) Pinpointed MIP spectrum, indicated by arrow in panel c. (c) MIP images of phosphatase activity profile in living SJSA-1 cells. Scale bar=30 μm (d) Quantification of MIP signal intensity of Phos-CN(S) and Phos-CN(P) in cells. n=10 cells. Data are presented as mean±SD. (e) MIP images of phosphatase activity profile in phosphatase inhibitor-pretreated SJSA-1 cells. Scale bar=30 μm (f) Quantification of MIP signal intensity of Phos-CN(S) and Phos-CN(P) in phosphatase inhibitor-pretreated SJSA-1 cells. n=10 cells. Data are presented as mean±SD. (g) Quantitation of product-to-substrate ratio ([P]/[S]) in the cells from PIC-pretreated and PIC-free groups. n=11 cells. Data are presented as mean±SD. (h) Confocal fluorescence image of SJSA-1 cells incubated with NBD-label phosphatase activity reporter (NBD-Phos-CN(S), 50 μM, 1h). Scale bar=30 μm (i) Comparison of SNR between MIP imaging of C≡N (at 2174 cm−1) and fluorescence imaging of NBD in cancer cells. Each dot represents a cell. n=10 cells. Data are presented as mean±SD. (j) Time-course MIP images of the phosphatase activity as Phos-CN(S) (50 μM in PBS, room temperature) was added to cells. Scale bar=30 μm (k) Pinpoint MIP spectrum, indicated by arrow in panel l (l) MIP images of caspase 3/7 activity profile in Doxorubicin-pretreated SJSA-1 cells. Scale bar=30 μm (m) Quantification of MIP intensity of Casp-CN(S) and Casp-CN(P) in Dox-pretreated cells and the cells from Dox-free control group. n=10 cells. Data are presented as mean±SD.

Article Snippet: Caspase-3/7 inhibitor Z-DEVD-FMK was purchased from Sigma Aldrich.

Techniques: Activity Assay, Imaging, Quantitation Assay, Fluorescence, Incubation, Comparison

Journal: Nature methods

Article Title: Mapping Enzyme Activity in Living Systems by Real-Time Mid-Infrared Photothermal Imaging of Nitrile Chameleons

doi: 10.1038/s41592-023-02137-x

Figure Lengend Snippet: (a) Simultaneous visualization of phosphatase and caspase-3/7 activity profile in Dox-pretreated SJSA-1 cells. Experiment was repeated independently 12 times with similar results. (b) Colocalization analysis of the mapping in (a). (c) Spatial interaction between phosphatase and caspase-3/7 in Dox-pretreated SJSA-1 cells. Similar results were observed in 12 independent experiments. (d) Intensity plot of Phos-CN(P) along the arrows in (c). (e) Correlation scatterplot of caspase-3/7 and phosphatase activity in Dox-pretreated SJSA-1 cells (f) Cell viability assay of SJSA-1 cells incubated with Dox in the presence and absence of phosphatase inhibitor cocktails. Phosphatase inhibitor cocktails were diluted by 4000 and 8000 times respectively using culture medium. n=3 independent experiments. Data are presented as mean±SD.

Article Snippet: Caspase-3/7 inhibitor Z-DEVD-FMK was purchased from Sigma Aldrich.

Techniques: Activity Assay, Viability Assay, Incubation