Journal: Molecular Cancer Therapeutics

Article Title: Silencing of Tubulin Binding Cofactor C Modifies Microtubule Dynamics and Cell Cycle Distribution and Enhances Sensitivity to Gemcitabine in Breast Cancer Cells

doi: 10.1158/1535-7163.mct-10-0568

Figure Lengend Snippet: Figure 1. Expression levels of different proteins in MCF7 cells with low TBCC levels. A, representative blots corresponding to TBCC, a-tubulin, b-tubulin, bIII-tubulin, TBCD, Arl2, and b-actin in MC-1, MC-2, MC-3, MP6.1, MP6.2, and MP6.3 cells. B, representative blots corresponding to a-tubulin and b-tubulin in NPT heterodimers, PT heterodimers, microtubule heterodimers, and total pool of tubulins in MC-1 and MP6.1 cells.

Article Snippet: MCF7 human mammary adenocarcinoma (American Type Culture Collection; no authentification was done by the authors) were cultured and transfected with pcDNA6/C or empty pcDNA6 as previously described (19).

Techniques: Expressing

Journal: Molecular Cancer Therapeutics

Article Title: Silencing of Tubulin Binding Cofactor C Modifies Microtubule Dynamics and Cell Cycle Distribution and Enhances Sensitivity to Gemcitabine in Breast Cancer Cells

doi: 10.1158/1535-7163.mct-10-0568

Figure Lengend Snippet: Figure 4. Cell cycle distribution and expression level of related proteins. A, percentages of cells in the S-phase and G2-M phase of the cell cycle. MC and MP6 represent the average value of 3 clones of MC or MP6, respectively. MP6.1 þ siRNA SCR, and TBCC correspond to MP6.1 cells transfected with siRNA scrambled and siRNA-targeting TBCC, respectively. Values differ significantly at P < 0.05 (*). Bars represent SD. B, representative blots showing TBCC content in MCF7 cells 48 hours after double-transient transfection with siRNA-targeting TBCC and scrambled siRNA. C, representative blots corresponding to the expression levels of CDK1, cyclin B, CDK2, cyclin E, p21, and p53 at basal levels and after 24 hours exposure to gemcitabine (1 nmol/L) in MC-1 and MP6.1 cells. D, quantification of the expression levels (ratio of protein versus b-actin) of CDK1, cyclin B, CDK2, cyclin E, p21, and p53 in TBCC models.

Article Snippet: MCF7 human mammary adenocarcinoma (American Type Culture Collection; no authentification was done by the authors) were cultured and transfected with pcDNA6/C or empty pcDNA6 as previously described (19).

Techniques: Expressing, Clone Assay, Transfection

Journal: Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology

Article Title: The entomotoxin Jack Bean Urease changes cathepsin D activity in nymphs of the hematophagous insect Dipetalogaster maxima (Hemiptera: Reduviidae).

doi: 10.1016/j.cbpb.2020.110511

Figure Lengend Snippet: Fig. 1. DmCatD expression profile (mRNA and protein) in fifth instar nymphs of D. maxima. The anterior (AM) and posterior midgut (PM), the fat body (FB), the Hemocytes (He) and the hemolymph (Hemo) from fed nymphs were processed for qPCR (A) and Western blot (B), as described in Materials and Methods. For qPCR, total RNA was extracted and the DmCatD transcripts were quantified using specific primers and 18S ribosomal RNA as normalizer. ***P < 0.0001 vs fat body, hemocytes and posterior midgut (n = 5). For Western blot, approximately 50 μg of proteins were loaded in each lane, separated by 15% SDS-PAGE and probed with the anti-CatD antibody. Whole cell MCF7 lysate (0.2 μg) was employed as a positive control. Densitometric analyses of three independent blots were performed upon the intensity of the pro-DmCatD and DmCatD bands detected in each lane and the amount of total proteins loaded and stained with Ponceau S.

Article Snippet: Rabbit polyclonal anti-human cathepsin D antibody (sc-10725) and MCF7 whole cell lysates (Santa Cruz Biotechnology, Palo Alto, CA, USA); Alexa Fluor 568-conjugated goat anti-rabbit IgG antibody and 4′,6-diamidino-2-phenylindole (DAPI) (Molecular Probes, Carlsbad, CA, USA); Tissue-Tek embedding medium Optimal Cutting Temperature (OCT, Miles, Elkhart, IN, USA); Fluorsave (Calbiochem, Darmstadt, Germany); electrophoresis protein standards (New England Biolabs, Ipswich, MA, USA); MMLV reverse transcriptase (Promega, Heidelberg, Germany); PCR Primers (Sigma Genosys, Houston, TX, USA); MasterPure RNA Purification Kit (Epicenter Biotechnologies, Madison, WI, USA); Power SYBR® Green PCR Master Mix (Applied Biosystems, Foster City, CA, USA) were from indicated commercial sources.

Techniques: Expressing, Western Blot, SDS Page, Positive Control, Staining