rabbit polyclonal anti active caspase 3  (Cell Signaling Technology Inc)

Journal: Nature Communications

Article Title: Rosiglitazone and trametinib exhibit potent anti-tumor activity in a mouse model of muscle invasive bladder cancer

doi: 10.1038/s41467-024-50678-2

Figure Lengend Snippet: A , B Expression of activated Caspase-3 in tumors of mice exposed to BBN for 5 months then treated with vehicle for 7 days, image shown with A and without B Ecad. C , D Expression of activated Caspase-3 in 5 M BBN tumors treated with Rosi+Tram for 7 days, image shown with C and without D Ecad. E Bar graph showing the percentage of cells expressing activated caspase-3 in BBN-induced tumors treated for 7 days with vehicle ( n = 3) or Rosi+Tram ( n = 3). Data given as means ± SD, significance calculated using two-tailed Mann-Whitney test, ***p = 0.0001. F Bar graph showing the percentage of cells expressing Ki67 in BBN-treated mice after vehicle ( n = 3) or Rosi+Tram ( n = 3) for 7 days. Data given as means ± SD, significance calculated using two-tailed Mann-Whitney test, ****p < 0.0001. G Apoptosis assayed by AnnexinV-eFluor450/7-AAD double staining in BBN963 cells treated with DMSO, Rosi, Tram, or combined Rosi+Tram for 72 h. Cells undergoing early apoptosis were AnnexinV+/7-AAD: late apoptosis: AnnexinV+/7-AAD+. H Heatmap showing normalized expression of proapoptotic genes, caspases, and pro-survival genes in Rosi+Tram treated BBN963 cells compared to controls based on RNAseq analysis. I – L Decreased proliferation determined by Ki67 staining in BBN963 cells after Rosi ( n = 4) J , Tram ( n = 4) K , or combined Rosi+Tram ( n = 4) L compared to controls ( n = 4) I . M – P Down-regulation of Ccnd1 in BBN963 cells after treatment with Rosi N , Tram O , or Rosi+Tram P compared to controls M . Scale bars: 50 µm. Q Heatmap of normalized gene expression showing down-regulation of Ccnd1 and genes that positively regulate cell cycle progression and up-regulation of genes that inhibit progression in BBN963 cells treated with Rosi+Tram for 72 h compared to DMSO treated controls. R Bar graph showing number of Ccnd1-positive BBN963 cells after 72-h treatment with DMSO ( n = 4), Rosi ( n = 4), Tram ( n = 4), and combined Rosi+Tram ( n = 4). Data given as means ± SD, significance calculated using two-tailed unpaired Welch’s t-test, *** p = 0.0006, ****p < 0.0001. Source data are provided as a Source Data file. Cartoons in Fig. 3 created with BioRender.com released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license ( https://creativecommons.org/licenses/by-nc-nd/4.0/deed.en ).

Article Snippet: Rabbit Polyclonal Anti- Active Caspase-3 , Promega , Cat#G7481 , N/A , 1:300.

Techniques: Expressing, Two Tailed Test, MANN-WHITNEY, Double Staining, Staining

Journal: Nature Communications

Article Title: Rosiglitazone and trametinib exhibit potent anti-tumor activity in a mouse model of muscle invasive bladder cancer

doi: 10.1038/s41467-024-50678-2

Figure Lengend Snippet:

Article Snippet: Rabbit Polyclonal Anti- Active Caspase-3 , Promega , Cat#G7481 , N/A , 1:300.

Techniques:

Journal: Scientific Reports

Article Title: Spatial sequestration of activated-caspase 3 in aggresomes mediates resistance of neuroblastoma cell to bortezomib treatment

doi: 10.1038/s41598-024-54140-7

Figure Lengend Snippet: Casp3A accumulates in aggresomes of resistant CLB-Sedp cells. (A,B) 15 μg of whole protein lysates from CLB-Ga ( A ) and CLB-Sedp ( B ) cells treated or not with 10 nM or 10 μM bortezomib for 24 h were separated by SDS-PAGE electrophoresis and transferred onto nitrocellulose membranes. Detection of Casp3 and PARP-1 by anti-Casp3 mAb and anti-Parp1 mAb, respectively. Ku80 was used as a loading control. Original blots are included in supplementary Fig. . ( C ) For CLB-Sedp cells: quantification of Casp3 was conducted using Image J software. Histogram representing the Casp3 signal intensity normalized against Ku80 (n = 2 ± SEM). ( D ) SHEP, CLB-Sedp, CLB-Ga, CLB-Boult and SKNAS cells treated or not with 10 μM bortezomib for 24 h were analyzed by immunofluorescence (IF). Cells were fixed, permeabilized and stained for vimentin. The number of vimentin cages corresponding to aggresomes was counted by microscopic observation of 10 random fields. Histograms represent the percentage of cells containing aggresomes. ( E ) IF analysis by confocal microscopy of vimentin subcellular localization in CLB-Ga and CLB-Sedp cells treated or not with 10 μM or 10 nM bortezomib during 24 h. Fluorescence micrographs of vimentin cages (anti-vim antibody, red signal) on fixed and permeabilized CLB-Ga and CLB-Sedp cells. Hoechst dye was used to stain the nuclei (blue) (scale bar = 10 μm). ( F ) Quantification of vimentin cages by 10 random fields in CLB-Ga and CLB-Sedp cells. Histograms represent the normalized value of aggresome formation. ( G ) IF analysis by confocal microscopy of Casp3A and vimentin subcellular localization in CLB-Sedp cells treated or not with 10 nM or 10 μM bortezomib for 24 h. Fluorescence micrographs of Casp3A (anti-Casp3A antibody, red signal) and Vimentin (Anti-vimentin mAb, green signal) on fixed and permeabilized cells. Hoechst dye was used to stain the nuclei (blue) (scale bar = 20 μm). ( H ) Superposition of green and red signals is presented on enlarged views of the merged images (white arrow). For all panels NT: Not treated; MT: Mock treated.

Article Snippet: Detection was performed using anti-vimentin (M0725 DaKoCytomation) coupled to a PLA anti-mouse probe, and rabbit polyclonal antibody anti-activated Casp3 (9661S Cell Signaling) coupled to a PLA anti-rabbit probe.

Techniques: SDS Page, Electrophoresis, Control, Software, Immunofluorescence, IF-cells, Staining, Confocal Microscopy, Fluorescence