Journal: International Journal of Biological Sciences

Article Title: 11β-Hydroxysteroid Dehydrogenase Type 1 Facilitates Osteoporosis by Turning on Osteoclastogenesis through Hippo Signaling

doi: 10.7150/ijbs.82933

Figure Lengend Snippet: Antibodies used in this study

Article Snippet: Ctsk (WB) , Santa Cruz , sc-48353.

Techniques:

Journal: Molecular pharmaceutics

Article Title: Surface-Functionalized Stem Cell-Derived Extracellular Vesicles for Vascular Elastic Matrix Regenerative Repair.

doi: 10.1021/acs.molpharmaceut.2c00769

Figure Lengend Snippet: Figure 1. CatK inhibitory effect of CKBP sequences. (A) Inhibition of rat recombinant CatK enzyme using CKBP sequences of varying lengths, and the inhibitory effect was determined using a CatK inhibitory assay. (In all plots, values shown indicate the mean ± SE of measurements made in n = 6. * denotes p < 0.05 compared to treatment controls (TC), # denotes p < 0.05 compared to CKBP-25-S group); (B) Dose-dependent inhibition of rat-recombinant CatK activity using CKBP-25 sequences in varying concentrations, determined using a CatK inhibitory assay. (In all plots, values shown indicate the mean ± SE of measurements made in n = 6. * denotes p < 0.05 compared to TC). (C) Inhibition of CatK activity in EaRASMCs using CKBP-25 and CKBP-25-S sequences, determined by a CatK inhibitory assay. (In all plots, values shown indicate the mean ± SE of measurements made in n = 6. * denotes p < 0.05 compared to the TC, # denotes p < 0.05 compared to the CKBP-25-S group).

Article Snippet: CKBP inhibitory effects on active CatK enzyme sequences were quantified using a CatK activity assay kit (RayBiotech Inc., Peachtree Corners, GA).

Techniques: Inhibition, Recombinant, Activity Assay

Journal: Molecular pharmaceutics

Article Title: Surface-Functionalized Stem Cell-Derived Extracellular Vesicles for Vascular Elastic Matrix Regenerative Repair.

doi: 10.1021/acs.molpharmaceut.2c00769

Figure Lengend Snippet: Figure 8. CatK activity in EaRAMSC after treatment with EV/EV- CKBP. CatK activity within EaRAMSC after treatment with EV/EV- CKBP (50 μg/mL for 24 h) was evaluated using the CatK inhibitory assay kit. Cells were pretreated with cytokines (IL-1β+TNF-α; 10 ng/ mL for 3 h) before EV/EV-CKBP addition (except for NT groups). (In all plots, the values shown indicate the mean ± SE of measurements made with n = 6)

Article Snippet: CKBP inhibitory effects on active CatK enzyme sequences were quantified using a CatK activity assay kit (RayBiotech Inc., Peachtree Corners, GA).

Techniques: Activity Assay

Journal: Molecular pharmaceutics

Article Title: Surface-Functionalized Stem Cell-Derived Extracellular Vesicles for Vascular Elastic Matrix Regenerative Repair.

doi: 10.1021/acs.molpharmaceut.2c00769

Figure Lengend Snippet: Figure 6. CatK activity in EaRAMSC after cytokine treatment (CT). CatK activity in EaRASMC was determined using the CatK inhibitory assay, after treatment with cytokines (IL-1β + TNF-α) for varying time and concentration, and compared with untreated cells (NT). CatK activity in EaRAMSCs (CT 10 ng/mL−3 h treated group) was inhibited using an inhibitor supplied by the kit manufacturer. (In all plots, values shown indicate the mean ± SE of measurements made in n = 6. * denotes p < 0.05 compared to the controls (NT), # denotes p < 0.05 compared to CT (10 ng/mL−30 min) group).

Article Snippet: CKBP inhibitory effects on active CatK enzyme sequences were quantified using a CatK activity assay kit (RayBiotech Inc., Peachtree Corners, GA).

Techniques: Activity Assay, Concentration Assay

Journal: Molecular pharmaceutics

Article Title: Surface-Functionalized Stem Cell-Derived Extracellular Vesicles for Vascular Elastic Matrix Regenerative Repair.

doi: 10.1021/acs.molpharmaceut.2c00769

Figure Lengend Snippet: Figure 9. Immunofluorescence studies showing expression of key proteolytic/regenerative marker proteins after treatment with EV/EV-CKBP. (A) Representative fluorescent microscopic images showing expression of CatK, MMP2, and Lox in EaRASMC cultures after the EV, EV-CKBP treatment (50 μg/mL for 24 h). Cells were pretreated with cytokines (IL-1β + TNF-α; 10 ng/mL for 3 h) before EV/EV-CKBP addition in these experiments. Cell nuclei were stained with DAPI (blue), and the proteins of interest were visualized with a secondary antibody conjugated to Alexa Fluor 546 dye (red), and images were acquired using fluorescent microscopy at 20× magnification (Scale bar-132.6 μm) (B) Quantification of red fluorescent intensity using Image J software across the treatment groups. (In all plots, values shown indicate the mean ± SE of measurements made in n = 3, * denotes p < 0.05 compared to TC groups).

Article Snippet: CKBP inhibitory effects on active CatK enzyme sequences were quantified using a CatK activity assay kit (RayBiotech Inc., Peachtree Corners, GA).

Techniques: Immunofluorescence, Expressing, Marker, Staining, Microscopy, Software

Journal: Molecular pharmaceutics

Article Title: Surface-Functionalized Stem Cell-Derived Extracellular Vesicles for Vascular Elastic Matrix Regenerative Repair.

doi: 10.1021/acs.molpharmaceut.2c00769

Figure Lengend Snippet: Figure 12. Ex vivo binding of EV/EV-CKBP onto elastase pretreated artery segments. (A) Binding of an activatable CatK fluorescent probe onto elastase (PPE)-pretreated porcine aortic tissues. Representative images demonstrating background tissue auto-fluorescence from nontreated (NT) samples with CatK probe bound to untreated tissue segments (NT + CatK680) and PPE-treated tissue segments (PPE + CatK680) are shown in the images. (B) Quantification of total radiant efficiency from the region of interest for each treatment group. (In all plots, the values shown indicate the mean ± SE of measurements made in n = 3. * denotes p < 0.05 compared to controls (NT), # denotes p < 0.05 compared to NT + CatK680 group). (C) Binding of EV-Cy5 and EV-CKBP-Cy5 (100 μg/mL for 20 min) to PPE pretreated porcine aortic tissues. Representative images demonstrating background tissue auto-fluorescence from PPE-treated control (TC), EV, and EV-CKBP incubated tissue segments are shown in the images. (D) Quantification of total radiant efficiency from the region of interest for each treatment group. (In all plots, values shown indicate mean ± SE of measurements made in n = 3. * denotes p < 0.05 compared to controls (TC), # denotes p < 0.05 compared to the EV treated group)

Article Snippet: CKBP inhibitory effects on active CatK enzyme sequences were quantified using a CatK activity assay kit (RayBiotech Inc., Peachtree Corners, GA).

Techniques: Ex Vivo, Binding Assay, Fluorescence, Control, Incubation

Journal: PLoS ONE

Article Title: Cathepsin K Is Present in Invasive Oral Tongue Squamous Cell Carcinoma In Vivo and In Vitro

doi: 10.1371/journal.pone.0070925

Figure Lengend Snippet: The knockdown of cathepsin K mRNA has significantly decreased the invasion of HSC-3 shRNA CTSK cells in the myoma organotypic model when compared with HSC-3 shRNA control cells (A–B). HSC-3 shRNA CTSK cells showed a reduction in cathepsin K mRNA levels when compared with wild type cells and HSC-3 cells transfected with a control shRNA (scramble) *p<0.0001 (C). The invasion depth, invasion area, and invasion index were all significantly reduced for HSC-3-shRNA CTSK cells (*p = 0.0006, **p<0.0001, and ***p = 0.0008, respectively) (D–F). Scale bars 200 µm.

Article Snippet: CTSK shRNA and control shRNA lentiviral particles (reference numbers sc-29936-V and sc-108080) were purchased from Santa Cruz Biotechnology (USA).

Techniques: Knockdown, shRNA, Control, Transfection