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

Journal: Oncotarget

Article Title: The TGFα-EGFR-Akt signaling axis plays a role in enhancing proinflammatory chemokines in triple-negative breast cancer cells

doi: 10.18632/oncotarget.25389

Figure Lengend Snippet: ( A ) Heatmap for RNA expression levels of chemokines in representative human TNBC (BT549, MB231 and MB468) and non-TNBC (T47D, MCF7) cell lines. ( B ) Heatmap for RNA expression levels of chemokine receptors in TNBC and non-TNBC cell lines. After isolating total RNA and choosing the qualified RNAs, a human chemokine PCR array was performed. Red trend indicates high expression levels of chemokines. ( C ) The status of estrogen receptor (ER), progesterone receptor (PR) and HER2 expression in TNBC and LA-BC cells. ( D ) Average intensity for the expression levels of dominant chemokines and chemokine receptors in TNBC and LA-BC cells. Red and blue bars indicate TNBC and LA-BC cells, respectively.

Article Snippet: The PIK3CA activity in BT549 and MCF7 cells (lysate) after time-dependent treatment with EGF and TNF was analyzed by an ELISA kit (Aviva Systems Biology Corporation, San Diego, CA, USA) according to the manufacturer's instructions.

Techniques: RNA Expression, Expressing

Journal: Oncotarget

Article Title: The TGFα-EGFR-Akt signaling axis plays a role in enhancing proinflammatory chemokines in triple-negative breast cancer cells

doi: 10.18632/oncotarget.25389

Figure Lengend Snippet: ( A ) Heatmap for RNA expression levels of EGFR family members in human BC tissues from TCGA-based dataset using Gitools 2.3.1. ( B ) Statistical analysis for RNA expression levels of EGFR family members in human BC tissues. The red, yellow, blue and green colors indicate BL, HER2, LA and LB samples, respectively. The asterisk ( * ) and hash (#) indicate a statistically significant increase and decrease ( p ≤ 0.05) as calculated by ANOVA and Tukey's pairwise comparisons, respectively. ( C ) Heatmap for RNA expression levels of EGFR family members based on analysis of the GEO dataset (Accession: GSE12777) for 51 human BC cell lines using Gitools 2.3.1. Pink, yellow and green dots indicate high expression levels in BL-TNBC, HER2-BC and LB-BC cells, respectively. ( D ) Protein levels of EGFR family members in representative TNBC (MB468, MB231, and BT549) and non-TNBC (MCF7 and T47D) cells. β-actin was used as the loading control.

Article Snippet: The PIK3CA activity in BT549 and MCF7 cells (lysate) after time-dependent treatment with EGF and TNF was analyzed by an ELISA kit (Aviva Systems Biology Corporation, San Diego, CA, USA) according to the manufacturer's instructions.

Techniques: RNA Expression, Expressing, Control

Journal: Oncotarget

Article Title: The TGFα-EGFR-Akt signaling axis plays a role in enhancing proinflammatory chemokines in triple-negative breast cancer cells

doi: 10.18632/oncotarget.25389

Figure Lengend Snippet: Chemokine signatures in non-TNBC (MCF7) and TNBC (BT549) cells after a 1-h stimulation with recombinant human EGF (10 ng/ml) as revealed by a human chemokine PCR array. Chemokines with duplicate average cycle threshold of <30 are considered dominant. The asterisk ( * ) and hash ( # ) indicate increases and decreases that are larger than two-fold, respectively.

Article Snippet: The PIK3CA activity in BT549 and MCF7 cells (lysate) after time-dependent treatment with EGF and TNF was analyzed by an ELISA kit (Aviva Systems Biology Corporation, San Diego, CA, USA) according to the manufacturer's instructions.

Techniques: Recombinant

Journal: Oncotarget

Article Title: The TGFα-EGFR-Akt signaling axis plays a role in enhancing proinflammatory chemokines in triple-negative breast cancer cells

doi: 10.18632/oncotarget.25389

Figure Lengend Snippet: ( A ) Basal protein levels of Akt and Erk in TNBC (BT549, MB231, MB468) and non-TNBC (MCF, T47D) cell lines. ( B ) Basal protein levels for NF-κB family members p65 (Rel A), Rel B, NF-κB (100/52), and NF-κB (105/50) in TNBC and non-TNBC cells. Human SKOV-3 and OVCAR-3 ovarian cancer cells were used as positive controls for the expression of NF-κB signaling components. ( C ) Protein expression profiles of IκB, Akt, Erk and their phosphorylated forms in BT549 and MCF7 cells in response to EGF (10 ng/ml) and TNF (10 ng/ml) treatments. β-actin was used as the loading control.

Article Snippet: The PIK3CA activity in BT549 and MCF7 cells (lysate) after time-dependent treatment with EGF and TNF was analyzed by an ELISA kit (Aviva Systems Biology Corporation, San Diego, CA, USA) according to the manufacturer's instructions.

Techniques: Expressing, Control