ctsb  (Bioss)

Journal: Molecular Therapy. Nucleic Acids

Article Title: SARS-CoV-2 infection in hiPSC-derived neurons is cathepsin-dependent and causes differential accumulation of HIF1ɑ and phosphorylated tau

doi: 10.1016/j.omtn.2025.102726

Figure Lengend Snippet: Cathepsin inhibitors block SARS-CoV-2 infection in neurons (A) Effectiveness of 0.2 μM apilimod, 25 μM nafamostat, 3–30 μM CA-074-ME and 3–30 μM SB412515 in blocking neuronal infection by the Wuhan strain of SARS-CoV-2. For each drug treatment, values were normalized to the corresponding concentrations of vehicle (DMSO) controls. Analysis included 6 replicate wells in two independent batches. (B) Effectiveness of 0.2 μM apilimod, 25 μM nafamostat, 3–30 μM CA-074-ME and 3–30 μM SB412515 in blocking neuronal infection by the Omicron XBB.1.5 strain of SARS-CoV-2. N = 6 replicate wells. (C) Representative images of data shown at A. Scale bars 600 μm (main) and 100 μm (zoom-in). (D) mRNA expression of cathepsin B (CTSB) and L (CTSL) in hiPSC-derived neurons, astrocytes, microglia, and neuron-astrocyte co-cultures. Data shown as fold change to the housekeeping gene GAPDH . (E) Effectivity of 25 μM nafamostat, 0.5 μM apilimod, 3–22.5 μM SB41251, and 3–30 μM CA-074-ME in blocking SARS-CoV-2 infection in Vero E6 cells, (F) and A549-AT cells. The data are displayed as fold-change to DMSO (reference level). N = 6 replicate wells. (G) Cytotoxicity assay of 3–22.5 μM SB41251 and 3–30 μM CA-074-ME in Vero E6 and A549-AT cell lines. DMSO was used as negative and 9 μM UCN-01 as a positive control. N = 8 replicate wells. All statistical significances were assessed using Kruskal-Wallis test with Dunn’s correction for multiple comparison. All comparisons were made to the SARS-CoV-2 infected DMSO control. Only significant results are denoted. ∗, p ≤ 0.05; ∗∗, p ≤ 0.01; ∗∗∗, p ≤ 0.001; ∗∗∗∗, p ≤ 0.0001. In all the graphs, the mean is indicated and the error bars are the standard deviation.

Article Snippet: We ran RT-qPCR using a Maxima probe/ROX qPCR master mix and the following TaqMan primers: ACE2 (HS01085333_m1), cathepsin B (Hs00947439_m1), cathepsin L (Hs00964650_m1), and GAPDH (Hs99999905_m1) (ThermoFisher Scientific) on a Bio-Rad CFX96 real-time system.

Techniques: Blocking Assay, Infection, Expressing, Derivative Assay, Cytotoxicity Assay, Positive Control, Comparison, Control, Standard Deviation

Journal: Journal of Pain Research

Article Title: Efficacy and Potential Mechanisms of Yi-Ping Lin Circum-Knee Acupuncture in the Treatment of Knee Osteoarthritis

doi: 10.2147/JPR.S547968

Figure Lengend Snippet: Protein interactions analysis and validation of hub genes. ( A – C ) General diagram of protein interactions analysis encoded by three groups of DEGs ( A ), Cluster 1 ( B ), and Cluster 2 ( C ) obtained according to the MCODE algorithm. ( D – H ) RT-qPCR was used to analyze the mRNA expression levels of hub genes Bgn ( D ), Ctsb ( E ), Lum ( F ), Serpine1 ( G ), and Tnfaip6 ( H ). ( I – N ) Western blot ( I ) was used to verify the protein expression levels of hub genes Bgn ( J ), Ctsb ( K ), Lum ( L ), Serpine1 ( M ), and Tnfaip6 ( N ). Statistical significance was indicated as * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001.

Article Snippet: Antibodies: mouse anti-β-actin monoclonal antibody (1:4000, TA-09, Zhongsui Jinqiao, China), Bgn (1:2000, 16409-1-AP, proteintech, China), Serpine1 (1:2000, bs-1704R, Bioss, China), Ctsb (1:2000, bs-1500R, Bioss, China), Tnfaip6 (1:1000, 13321-1-AP, proteintech, China), Lum (1:1000, 10677-1-AP, proteintech, China), Goat Anti Mouse IgG -HRP (1:4000, M21001L, Abmart, Shanghai, China), Goat Anti Rabbit IgG-HRP (1:4000, M21002L, Abmart, Shanghai, China).

Techniques: Biomarker Discovery, Quantitative RT-PCR, Expressing, Western Blot