rabbit anti ki67 ab  (Novus Biologicals)

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: A nanoencapsulated oral formulation of fenretinide promotes local and metastatic breast cancer dormancy in HER2/neu transgenic mice

doi: 10.1186/s13046-024-03213-6

Figure Lengend Snippet: Proliferative activity of Bio-nFeR-treated human and murine BC cells. (A) Flow cytometry analysis for the frequency of Ki67-positive cells in MCF7, MDA-MB-231 and Tubo lines. Cells were treated with 30µM, 80µM, 25µM Bio-nFeR, respectively, for 24 h. (B) Left: Representative confocal images of Ki67-positive nuclei of BC cell lines treated as in A; Right: Proliferation index was calculated as the ratio Ki67-positive/ total nuclei by automated pixel counting, see Methods. Scale bar 100 μm

Article Snippet: Anti-β-tubulin and anti-β-actin antibodies were purchased from Sigma-Aldrich; Phospho-S6 Ribosomal Protein (Ser240/244) #2215 and S6 Ribosomal Protein (5G10) Rabbit Ab #2217, Caspase-3 Rabbit Ab #9662, Caspase-7 (D2Q3L) Rabbit Ab #12,827, Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP ® Rabbit Ab #4511, p38 MAPK (D13E1) XP ® Rabbit Ab #8690, mTOR Rabbit Ab #2972, Phospho-mTOR (Ser2448) Rabbit Ab #2971, 4E-BP1 (53H11) Rabbit Ab #9644, Phospho-4E-BP1 (Ser65) Rabbit Ab #9451, Bcl-2 (D17C4) Rabbit Ab #3498, ALDH1 (D4R9V) Rabbit Ab #12,035, and Ki67 (D3B5) Rabbit Ab (IHC Formulated) #12,202, Ki67 (D3B5) Rabbit Ab #9129, PCNA (D3H8P) Rabbit Ab #13,110 were from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Activity Assay, Flow Cytometry

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: A nanoencapsulated oral formulation of fenretinide promotes local and metastatic breast cancer dormancy in HER2/neu transgenic mice

doi: 10.1186/s13046-024-03213-6

Figure Lengend Snippet: In vivo test of Bio-nFer on neuT mice. Bio-nFeR was administered at 100 mg/kg daily, 5 days/week, from week 14 to week 32 PB by oral gavage. (A) Time-course of tumor development in the mammary glands over time, post first tumor occurrence in vehicle (black) versus Bio-nFeR (red) mice. (B) Time to reach 10/10 mammary gland invasion in vehicle (black) versus Bio-nFeR (red) mice, post birth. (C) Average tumor volume at sacrifice in vehicle (black) versus Bio-nFeR (red) mice. (D) Scatter plot of average tumor burden (volume) at sacrifice in Bio-nFeR versus control mice. (E) Average tumor weight at sacrifice in vehicle (black) versus Bio-nFeR (red) mice. (F) Scatter plot of average tumor burden (weight) at sacrifice in vehicle versus Bio-nFeR mice. (G) Left: representative images of immunofluorescence staining of mammary tumors harvested ex vivo from vehicle-treated and Bio-nFeR-treated mice (20x, 0,7x zoom magnification, scale bar 50 μm). Ki67 (pseudocolored in green) and DAPI nuclear staining (blue); Right: proliferation index by Ki67 nuclear staining on sections. Data in A represent Mean ± SEM, *** P < 0.001 by paired Student’s t test with Wilcoxon test. Data in B were analyzed by Long-rank (Mantel-Cox) test, * P < 0.05 and. Data in C , D , E , F represent Mean ± SEM, * P < 0.05 and ** P < 0.01 by unpaired Student’s t test with Welch correction. Data in G represent Mean ± SD, * P < 0.05 by unpaired Student’s t test with Welch’s correction

Article Snippet: Anti-β-tubulin and anti-β-actin antibodies were purchased from Sigma-Aldrich; Phospho-S6 Ribosomal Protein (Ser240/244) #2215 and S6 Ribosomal Protein (5G10) Rabbit Ab #2217, Caspase-3 Rabbit Ab #9662, Caspase-7 (D2Q3L) Rabbit Ab #12,827, Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP ® Rabbit Ab #4511, p38 MAPK (D13E1) XP ® Rabbit Ab #8690, mTOR Rabbit Ab #2972, Phospho-mTOR (Ser2448) Rabbit Ab #2971, 4E-BP1 (53H11) Rabbit Ab #9644, Phospho-4E-BP1 (Ser65) Rabbit Ab #9451, Bcl-2 (D17C4) Rabbit Ab #3498, ALDH1 (D4R9V) Rabbit Ab #12,035, and Ki67 (D3B5) Rabbit Ab (IHC Formulated) #12,202, Ki67 (D3B5) Rabbit Ab #9129, PCNA (D3H8P) Rabbit Ab #13,110 were from Cell Signaling Technology (Danvers, MA, USA).

Techniques: In Vivo, Control, Immunofluorescence, Staining, Ex Vivo

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: A nanoencapsulated oral formulation of fenretinide promotes local and metastatic breast cancer dormancy in HER2/neu transgenic mice

doi: 10.1186/s13046-024-03213-6

Figure Lengend Snippet: Immunohistochemistry analysis of Ki67 and PCNA on lung metastasis from Bio-nFeR versus control mice. (A) Left: Representative IHC images of Ki67-positive nuclei on lung metastasis from Bio-nFeR versus vehicle mice; Right: Proliferation index by Ki67 staining in treated and control metastases; (B) Left: Representative IHC images of PCNA-positive nuclei in lung metastases from Bio-nFeR versus vehicle mice; Right: Proliferation index by PCNA nuclear staining in treated and control metastases. Quantifications were obtained by analyzing images at 20x magnification. Proliferation index was calculated by automated pixel counting, as the ratio between Ki67-or PCNA-positive/total nuclei (see Methods and Additional file: Fig. ). Magnifications 5x and 20x, scale bares 200 and 50 μm, respectively

Article Snippet: Anti-β-tubulin and anti-β-actin antibodies were purchased from Sigma-Aldrich; Phospho-S6 Ribosomal Protein (Ser240/244) #2215 and S6 Ribosomal Protein (5G10) Rabbit Ab #2217, Caspase-3 Rabbit Ab #9662, Caspase-7 (D2Q3L) Rabbit Ab #12,827, Phospho-p38 MAPK (Thr180/Tyr182) (D3F9) XP ® Rabbit Ab #4511, p38 MAPK (D13E1) XP ® Rabbit Ab #8690, mTOR Rabbit Ab #2972, Phospho-mTOR (Ser2448) Rabbit Ab #2971, 4E-BP1 (53H11) Rabbit Ab #9644, Phospho-4E-BP1 (Ser65) Rabbit Ab #9451, Bcl-2 (D17C4) Rabbit Ab #3498, ALDH1 (D4R9V) Rabbit Ab #12,035, and Ki67 (D3B5) Rabbit Ab (IHC Formulated) #12,202, Ki67 (D3B5) Rabbit Ab #9129, PCNA (D3H8P) Rabbit Ab #13,110 were from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Immunohistochemistry, Control, Staining