cathepsin g inhibitor i  (MedChemExpress)

Journal: Cellular and Molecular Immunology

Article Title: Oncolytic virus M1 reinvigorates CD8 + T-cell immunity against glioblastoma through B-cell-dependent antigen cross-presentation in the spleen

doi: 10.1038/s41423-026-01396-w

Figure Lengend Snippet: B cells activate CD8 + T cells via antigen cross-presentation following OVM administration. Bubble plots of KEGG ( A ) and GO ( B ) pathway enrichment for DEGs in B cells following OVM versus vehicle treatment. C GSEA of B cells showing upregulation of the antigen processing and presentation pathway (GO:0019882) in B OVM cells compared with B veh cells. D Heatmap showing the average expression of MHC-related transcription factors and molecules, antigen processing and presentation, and cytokines and chemokines in B veh and B OVM cells. E Flow cytometric analysis of surface H-2Kb-SIINFEKL expression on B cells, with the results shown as the MFI. F – H B veh and B OVM cells were isolated from the spleens of GL261-OVA tumor-bearing mice treated with vehicle or OVM and cocultured with OT-1 CD8 + T cells ( F ). OT-1 CD8 + T-cell proliferation was measured by CFSE dilution ( G ), and activation was assessed by the expression of CD44 ( H ). I B veh and B OVM cells were incubated with OVA-Cy5 (10 µg/mL) for 24 h, and the proportion of OVA + B cells was determined. J Heatmap showing the average expression of lysosomal protease- and proteasome-associated genes in B veh and B OVM cells. B veh and B OVM cells were pretreated with the proteasome inhibitor lactacystin (2.5 µg/mL), the lysosome inhibitor cathepsin inhibitor I (10 µM) ( K ), or the ERAD inhibitor eeyarestatin I (3 µM) ( L ) before being cocultured with CD8 + T cells from tumor-bearing mice. CD8 + T-cell proliferation and the frequencies of IFN-γ + and GZMB + CD8 + T cells were then assessed via flow cytometry

Article Snippet: For inhibitor studies targeting the immunological synapse, proteasome, lysosome, or ER-associated degradation (ERAD) pathways, B cells were pretreated for 24 h with vehicle (DMSO), the ICAM1–LFA interaction inhibitor A-286982 (Selleck, S3408) (1 μM), the proteasome inhibitor lactacystin (MCE, HY-16594) (2.5 μg/mL), the lysosome inhibitor cathepsin inhibitor I (TargetMol, 225120--65--0) (10 μM), or the ERAD inhibitor Eeyarestatin I (TargetMol, 412960--54--4) (3 μM).

Techniques: Expressing, Isolation, Activation Assay, Incubation, Flow Cytometry

Journal: Redox Biology

Article Title: Lactate-driven ATP6V1B2 lactylation triggers asthmatic inflammation by linking lysosomal dysfunction to mitochondrial ROS-dependent pyroptosis

doi: 10.1016/j.redox.2026.104059

Figure Lengend Snippet: Effect of inhibiting Cathepsin B on mitochondrial dysfunction and tBid-Bax pathway. (A) Fluorescence imaging of mitochondrial membrane potential (JC-1) after administration of Cathepsin B inhibitor (Ca-074-Me). Scale bar: 100 μm. The ratio of red/green fluorescence intensity was quantified. (B) MitoSOX Red staining detects mitochondrial ROS. Scale bar: 100 μm. Quantification of MitoSOX Red fluorescence intensity is shown. (C) TEM images of mitochondrial ultrastructure. Scale bar: 2.0 μm. (D) Immunoblot analysis of Cathepsin B distribution in subcellular fractions (lysosome, cytosol, mitochondria). (E) Immunofluorescence staining of Cathepsin B (green) and LAMP1 (red). Scale bar: 100 μm. (F) Immunofluorescence staining of Cathepsin B (green) and MitoTracker (red). Scale bar: 40 μm. (G) Immunoblot analysis of Bid, tBid, XIAP, Bcl-2, Bcl-xL, and Bax protein expression. Data are expressed as Mean ± SEM. (n = 3 independent experiments) ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001; ns, not significant.

Article Snippet: In mechanistic studies, cells were pre-incubated with the following specific inhibitors for 2 h before stimulation: Ferrostatin-1, Necrosulfonamide, Z-VAD-FMK (10 μM), Z-IETD-FMK (10 μM), Z-DEVD-FMK (10 μM), Bafilomycin A1 (100 nM), Chloroquine (10 μM), 3-Methyladenine (3-MA, 3 mM), Cathepsin B inhibitor CA-074 Methyl Ester (Cat# HY-100350, MCE), BID inhibitor BI-6C9 (Cat# HY-103661, MCE), and calcium chelator BAPTA-AM (10 μM, Cat# HY-100545, MCE).

Techniques: Fluorescence, Imaging, Membrane, Staining, Western Blot, Immunofluorescence, Expressing

Journal: The Journal of Clinical Investigation

Article Title: Galectin-3 mediates lysosome-related inflammation within monocyte-derived macrophages in a mouse model of ischemic brain injury

doi: 10.1172/JCI194139

Figure Lengend Snippet: ( A ) Representative immunostaining of cathepsin B/GAL3/CD68 in WT/WT and WT/GAL3 –/– chimera mice 3 d after tMCAO. Scale bar: 50 μm. ( B ) Quantification of cathepsin B + staining areas within CD68 + areas of macrophages. ( C ) Quantification of CD68 + areas. N = 7/group. ( D ) MAP2 staining of brain in vehicle- or CA-074–treated (10 mg/kg, i.v., 2 h after tMCAO and then daily for 2 d) WT/WT or WT/GAL3 –/– chimeric mice 3 d after tMCAO. ( E ) Quantification of brain infarct volume. N = 5–6/group. ( F ) Cathepsin B activity was analyzed using the Magic Red cathepsin kit in GAL3-KO or WT BMDMs with vehicle or TD139 (10 μM) treatment. Scale bar: 50 μm. ( G ) Quantification of the percentages of Magic Red + cells among total cells. N = 6/condition. ( H ) Representative Magic Red staining in WT and GAL3-KO BMDMs treated with or without WT or GAL3-KO lysate for 6 h. Scale bars: 50 μm. ( I ) Quantification of the percentages of Magic Red + cells among total cells. N = 6/condition. Gray blocks show SD. *** P < 0.001 versus (WT BMDM + WT lysate); # P < 0.05, ## P < 0.01 versus (KO BMDM + WT lysate); $ P < 0.05 versus (WT BMDM + KO lysate). ( J ) Immunoblot and quantification of cathepsin B expression in cytosol of BMDMs under the indicated conditions for 6 h. N = 3/condition. ( K ) Coimmunostaining of LAMP2 and cathepsin B in BMDMs under specified conditions for 6 h. Scale bars: 5 μm, 2 μm (zoom). ( L ) Fluorescence intensity profiles of cathepsin B and LAMP2. ( M ) Pearson’s correlation analyses of LAMP2 and cathepsin B. N = 7/condition. ( N ) BMDMs in inserts treated with brain lysates and CA-074 (10 μM) or vehicle were cocultured with OGD neurons. Coverage areas of MAP2-stained neurons were quantified. N = 6/condition. Scale bar: 40 μm. * P < 0.05, ** P < 0.01, *** P < 0.001. Two-tailed, unpaired Student’s t test ( B , C , and E ) or 1-way ( G , J , M , and N ) or 2-way ( I ) ANOVA and Bonferroni’s test.

Article Snippet: Stock solutions of TD139, a specific GAL3 inhibitor (TargetMol) and CA-074 methyl ester, a specific cathepsin B inhibitor (MedChem Express) were prepared at concentrations of 10 mM for in vitro use and 10 mg/mL for in vivo use.

Techniques: Immunostaining, Staining, Activity Assay, Western Blot, Expressing, Fluorescence, Two Tailed Test