Journal: Cancer Prevention Research

Article Title: 3,3′-Diindolylmethane Enhances the Efficacy of Butyrate in Colon Cancer Prevention through Down-Regulation of Survivin

doi: 10.1158/1940-6207.capr-08-0142

Figure Lengend Snippet: Fig. 2. DIM down-regulates survivin in HT-29 cells. A, DIM down-regulated survivin in HT-29 cells. HT-29 cells were treated with various doses of DIM for 24 h. Survivin and β-tubulin protein levels were detected by Western blotting. B, DIM inhibited transcription of survivin mRNA. HT-29 cells were treated with DMSO or 40 μmol/L DIM for 24 h. Total RNA was isolated from cells and analyzed by real-time PCR as described in Materials and Methods. C, MG-132 blocked DIM-induced down-regulation of survivin. HT-29 cells were treated with DMSO, 40 μmol/L DIM, or 40 μmol/L DIM plus 2 μmol/L MG-132 for 24 h. Survivin and β-tubulin protein levels were detected by Western blotting. D, HT-29 cells were treated with DMSO, 40 μmol/L DIM for 24 h, 20 μmol/L cycloheximide for 2 h, or 40 μmol/L DIM for 22 h plus 20 μmol/L cycloheximide for additional 2 h. Survivin and β-tubulin protein levels were detected by Western blotting. E, DIM down-regulated cyclin B1. HT-29 cells were treated with various doses of DIM for 24 h. Cyclin B1, cdc2, and tubulin protein levels were detected by Western blotting. F, DIM inhibited survivin Thr34 phosphorylation. HT-29 cells were transfected with pCMV-Tag2A-survivin plasmids. Twenty-four hours after transfection, cells were treated with 40 μmol/L DIM together with 10 μmol/L MG-132 (to prevent protein degradation) for an additional 24 h. p-Thr34-survivin and tubulin levels were detected by Western blotting. Relative protein levels were quantified and shown under the gels. All experiments were repeated thrice.

Article Snippet: Anti–phospho-survivin (Thr34) polyclonal antibody was purchased from Novus.

Techniques: Western Blot, Isolation, Real-time Polymerase Chain Reaction, Phospho-proteomics, Transfection

Journal: Cell Death & Disease

Article Title: PLK1 and AURKB phosphorylate survivin differentially to affect proliferation in racially distinct triple-negative breast cancer

doi: 10.1038/s41419-022-05539-5

Figure Lengend Snippet: List of reagents and antibodies.

Article Snippet: 9 , Anti-p-survivin (S20) antibody , 1:1000 (WB) 1:100 (IHC-P) , NB11092717 , Novus Biologicals.

Techniques: Concentration Assay, cDNA Synthesis, SYBR Green Assay, Protease Inhibitor, Saline, Plasmid Preparation, Magnetic Beads, Membrane

Journal: Cell Death & Disease

Article Title: PLK1 and AURKB phosphorylate survivin differentially to affect proliferation in racially distinct triple-negative breast cancer

doi: 10.1038/s41419-022-05539-5

Figure Lengend Snippet: List of reagents and antibodies.

Article Snippet: 10 , Anti-p-survivin (T117) antibody , 1:1000 (WB) 1:100 (IHC-P) , MBS003339 , MyBioSource.

Techniques: Concentration Assay, cDNA Synthesis, SYBR Green Assay, Protease Inhibitor, Saline, Plasmid Preparation, Magnetic Beads, Membrane

Journal: Cell Death & Disease

Article Title: PLK1 and AURKB phosphorylate survivin differentially to affect proliferation in racially distinct triple-negative breast cancer

doi: 10.1038/s41419-022-05539-5

Figure Lengend Snippet: A BIRC5 expression level in AA and EA patients with TNBC (TCGA dataset). B Relative BIRC5 expression in AA and EA TNBC cell lines from Neve et al. (2006) ( B ) and our in-house TNBC cell lines ( C ) ( n = 3 each). D Immunoblot showing survivin levels in AA ( n = 3) and EA ( n = 3) TNBC cell lines. E – G Representative IHC images ( E ) and bar graphs ( F , G ) showing survivin H scores in AA and EA patients with TNBC in the Dekalb ( F ) and Emory ( G ) cohorts. H IF images showing the localization of survivin (green) in AA and EA TNBC cell lines. Nuclei were stained with Hoechst (blue) and tubulin (red). I Immunoblot showing p-survivin (S20, T117) levels in AA and EA TNBC cell lines ( n = 3 each). FPKM fragments per kilobase of transcript per million mapped reads, TPM transcripts per million mapped reads. Bars indicate mean ± SEM. Unpaired two-tailed Student’s t -test with Welch’s correction was used to determine statistical significance (* P < 0.05, *** P < 0.0005, ns = non-significant). Scale bars in E and H are 100 µm and 10 µm, respectively.

Article Snippet: 10 , Anti-p-survivin (T117) antibody , 1:1000 (WB) 1:100 (IHC-P) , MBS003339 , MyBioSource.

Techniques: Expressing, Western Blot, Staining, Two Tailed Test

Journal: Cell Death & Disease

Article Title: PLK1 and AURKB phosphorylate survivin differentially to affect proliferation in racially distinct triple-negative breast cancer

doi: 10.1038/s41419-022-05539-5

Figure Lengend Snippet: A , B Immunoblots showing the levels of PLK1, survivin, p-survivin (S20), and β-actin after PLK1 silencing ( A , B ) or inhibition ( C ) in AA and EA TNBC cell lines. D – F Immunoblots showing the levels of AURKB, survivin, p-survivin (T117), and β-actin after AURKB silencing ( D , E ), or inhibition ( F ) in AA and EA TNBC cell lines.

Article Snippet: 10 , Anti-p-survivin (T117) antibody , 1:1000 (WB) 1:100 (IHC-P) , MBS003339 , MyBioSource.

Techniques: Western Blot, Inhibition

Journal: Cell Death & Disease

Article Title: PLK1 and AURKB phosphorylate survivin differentially to affect proliferation in racially distinct triple-negative breast cancer

doi: 10.1038/s41419-022-05539-5

Figure Lengend Snippet: A – F Representative IHC images ( A , B ) and bar graphs ( C – F ) showing Ki-67 and survivin levels in AA ( C , E ) and EA ( D , F ) TNBC xenografts under various treatment conditions. G , H Immunoblots showing the levels of p-survivin (T117), p-survivin (S20), total survivin, AURKB, PLK1, and β-actin in AA ( G ) and EA ( H ) fresh-frozen xenograft tumor lysates from mice treated with volasertib, barasertib, or their combination ( n = 12 per treatment group). Bars represent mean ± SEM. Unpaired two-tailed Student’s t -test with Welch’s correction was used to determine statistical significance (**** P < 0.00005, ns = non-significant). The scale bar represents 100 µm.

Article Snippet: 10 , Anti-p-survivin (T117) antibody , 1:1000 (WB) 1:100 (IHC-P) , MBS003339 , MyBioSource.

Techniques: Western Blot, Two Tailed Test