Journal: Veterinary World

Article Title: Isolation, genetic characterization, and virulence profiling of different Aeromonas species recovered from moribund hybrid catfish ( Clarias spp.)

doi: 10.14202/vetworld.2023.1974-1984

Figure Lengend Snippet: Identity of Aeromonas spp. bacteria isolated from diseased catfish.

Article Snippet: Av-01, Av-02, Av-03, and Av-04 isolates were grouped into the clade of A . veronii bv veronii ATCC 35624 and Ad-01, Ad-02, and Ad-03 isolates into the clade of A. dhakensis P21 ( ).

Techniques: Bacteria, Isolation

Journal: International Journal of Biological Sciences

Article Title: A Novel Aurora-A Inhibitor, BPR1K0609S1, Sensitizes Colorectal Tumor cells to 5-Fluorofracil (5-FU) Treatment

doi: 10.7150/ijbs.5806

Figure Lengend Snippet: The effects of BPR1K0609S1 (BP) on cell cycle. HCT116 and their isogenic variants, HCT116 p53(-), HCT116 Puma(-), HCT116 Bax(-) HCT116 p21(-) and HCT116 Chk2(-), were treated with BP (800nM) for 48 h, and cell cycle profiles were determined by propidium iodide (PI) staining. Results were obtained from two independent experiments.

Article Snippet: HCT116 was purchased from ATCC and isogenic HCT116 variants deficient for p53, Puma, Bax, Chk2 or p21 were kindly obtained from Dr. Bert Vogelstein (Johns Hopkins University, Ref. 14-17).

Techniques: Staining

Journal: BMC Cancer

Article Title: Increased diacylglycerol kinase ζ expression in human metastatic colon cancer cells augments Rho GTPase activity and contributes to enhanced invasion

doi: 10.1186/1471-2407-14-208

Figure Lengend Snippet: Increased PAK1 expression and phosphorylation in SW620 cells. (A) Detergent extracts prepared from SW480 and SW620 cells were analyzed by immunoblotting for phosphorylated (p)PAK1, total PAK1 and tubulin. (B) Graph showing the quantification of total PAK1 levels by densitometric analysis of immunoblots. (C) Quantification of pPAK1 levels. The intensity of the upper pPAK1 band (arrow in A) was measured and normalized to the level of tubulin. Values are the average ± S.E.M. of four independent experiments. One asterisk denotes a significant difference (P < 0.05) and two asterisks, a highly significant difference (P < 0.01) from SW480 cells by Student’s t -test.

Article Snippet: ATCC: American Type Culture Collection; DAG: Diacylglycerol; DGK: Diacylglycerol kinase; DMEM: Dulbecco’s Modified Eagle Medium; FBS: Fetal bovine serum; PA: Phosphatidic acid; PAK1: p21-activated kinase 1.

Techniques: Expressing, Western Blot

Journal: Molecular Cancer

Article Title: Anti-tumor effects of fibroblast growth factor-binding protein (FGF-BP) knockdown in colon carcinoma

doi: 10.1186/1476-4598-10-144

Figure Lengend Snippet: Effects of FGF-BP expression on cell proliferation and colony formation . Clonal or mass-transfected shRNA expressing LS174T cell lines with reduced FGF-BP levels show decreased anchorage-dependent cell proliferation as determined in WST-1 assays (A). Anti-proliferative effects in LS174T cells are dependent on the extent of RNAi-mediated FGF-BP knockdown, and the correlation between residual FGF-BP levels and the proliferative activity establishes an FGF-BP 'gene-dose effect' on cell proliferation (B). Soft-agar assays reveal marked reductions of anchorage-independent proliferation upon FGF-BP knockdown in LS-174T (C) and in HT29 cells (D). Anti-proliferative effects are confirmed in HCT-116 cells that show decreased anchorage-dependent (E, left) and anchorage-independent proliferation (E, right) upon FGF-BP knockdown.

Article Snippet: LS174T, HCT-116 and HT29 colon carcinoma cells were obtained from the American Type Culture Collection (ATCC, Manassas, VA), HCT-116 p21 -/- were obtained from Dr. Bert Vogelstein [ ], and stable FGF-BP expressing SW-13 adrenal carcinoma cells have been described previously [ ].

Techniques: Expressing, Transfection, shRNA, Activity Assay

Journal: Molecular Cancer

Article Title: Anti-tumor effects of fibroblast growth factor-binding protein (FGF-BP) knockdown in colon carcinoma

doi: 10.1186/1476-4598-10-144

Figure Lengend Snippet: Effects of FGF-BP expression on cell cycle and apoptosis . Stable FGF-BP knockdown cells vs. control LS174T cells were treated with nocodazole 20 h prior to propidium iodide/FACS cell cycle analysis. Decreased percentages of cells in G2/M arrest demonstrate slower cell cycle progression after FGF-BP knockdown (A), while sub-G0 levels are increased (A; see also right bar diagram). Caspase-3/7 assays show the induction of apoptosis, represented by increased caspase-3/7 activity, upon FGF-BP knockdown in LS174 (B, left) or HCT-116 cells (B, right). Expression of exogenous FGF-BP leads to anti-apoptotic effects in endogenously FGF-BP-negative SW13 cells (C, right), but not in LS174T cells with already high endogenous FGF-BP levels (C, left).

Article Snippet: LS174T, HCT-116 and HT29 colon carcinoma cells were obtained from the American Type Culture Collection (ATCC, Manassas, VA), HCT-116 p21 -/- were obtained from Dr. Bert Vogelstein [ ], and stable FGF-BP expressing SW-13 adrenal carcinoma cells have been described previously [ ].

Techniques: Expressing, Cell Cycle Assay, Activity Assay

Journal: Molecular Cancer

Article Title: Anti-tumor effects of fibroblast growth factor-binding protein (FGF-BP) knockdown in colon carcinoma

doi: 10.1186/1476-4598-10-144

Figure Lengend Snippet: Analysis of cellular and molecular effects of FGF-BP knockdown . The effects of FGF-BP levels on cell proliferation are dependent on p21 as indicated by the absence of anti-proliferative effects of FGF-BP knockdown in HCT-116 p21 -/- cells (A). Dependence of antiproliferative GSK3β effects on FGF-BP. GSK3β inhibition by treatment of LS174T cells with 6-bromoindirubin-3-oxime (BIO) leads to the induction of proliferation (B, left). This effect is abrogated upon FGF-BP knockdown (B, right). Dependence of FGF2 cell proliferation-simulating effects on FGF-BP expression. Stimulation of LS174T cells by treatment with FGF2 leads to the induction of proliferation (C, left). This effect is largely abrogated upon FGF-BP knockdown (C, right). Anti-proliferative effects of transient siFGF-BP transfection (D).

Article Snippet: LS174T, HCT-116 and HT29 colon carcinoma cells were obtained from the American Type Culture Collection (ATCC, Manassas, VA), HCT-116 p21 -/- were obtained from Dr. Bert Vogelstein [ ], and stable FGF-BP expressing SW-13 adrenal carcinoma cells have been described previously [ ].

Techniques: Inhibition, Expressing, Transfection

Journal: Molecular Cancer

Article Title: p21 WAF1 expression induced by MEK/ERK pathway activation or inhibition correlates with growth arrest, myogenic differentiation and onco-phenotype reversal in rhabdomyosarcoma cells

doi: 10.1186/1476-4598-4-41

Figure Lengend Snippet: mRNA stabilization by TPA and transcriptional activation by U0126 of p21 WAF1 expression. (A). RD cells were transfected with plasmid expressing β-galactosidase (β-gal) gene and a plasmid carrying p21 promoter (DM-Luc). Luciferase activity was normalized for the expression levels of transfected β-gal protein. Data show mean values ± s.e.m. of triplicates of a representative experiment. (B) Northern blots from RD cells left untreated (C) or treated with TPA (upper panel) or U0126 (U) (lower panel) for 5 hours, pre-treated with 0.05 μg/ml of actinomycin D for 1 hour and then left untreated (ActD) or treated with TPA (ActD+TPA) or U0126 (ActD+U) for 5 hours. The levels of GAPDH mRNA are shown. Similar results were obtained in three independent experiments for A and two for B.

Article Snippet: The p21 WAF1 , MyoD and myogenin probes was obtained from the plasmid described below, while cyclin D1 probe was kindly provided by Dr. A. Arnold [ ] and GAPDH vector was provided by ATCC.

Techniques: Activation Assay, Expressing, Transfection, Plasmid Preparation, Luciferase, Activity Assay, Northern Blot

Journal: Molecular Cancer

Article Title: p21 WAF1 expression induced by MEK/ERK pathway activation or inhibition correlates with growth arrest, myogenic differentiation and onco-phenotype reversal in rhabdomyosarcoma cells

doi: 10.1186/1476-4598-4-41

Figure Lengend Snippet: MEK/ERK pathway sustains p21 WAF1 expression. (A) RD cells were left untreated (-) or treated with TPA in the absence or in the presence of U0126 for the times indicated. (B) RD cells were transfected with the constitutively active form of HA-tagged-MEK1 (ca MEK1), -MEK2 (ca MEK2) or with the empty vector (CMV). (C) RD cells were transfected with control siRNA (siRNA-C) or ERK1 and ERK2 siRNAs (siRNAERK1-2) and then left untreated (-) or treated with TPA (+) for 4 days. Immunoblots of total lysates were performed using specific antibodies capable of recognising p21 WAF1 , total and phosphorylated ERK1/2, hemagglutinin (HA) and α-tubulin. The data shown are representative of three independent experiments for A and two for B and C.

Article Snippet: The p21 WAF1 , MyoD and myogenin probes was obtained from the plasmid described below, while cyclin D1 probe was kindly provided by Dr. A. Arnold [ ] and GAPDH vector was provided by ATCC.

Techniques: Expressing, Transfection, Plasmid Preparation, Western Blot

Journal: Molecular Cancer

Article Title: p21 WAF1 expression induced by MEK/ERK pathway activation or inhibition correlates with growth arrest, myogenic differentiation and onco-phenotype reversal in rhabdomyosarcoma cells

doi: 10.1186/1476-4598-4-41

Figure Lengend Snippet: Effects of p38 inhibition on p21 WAF1 , myogenin and MyoD expression. (A and B) RD cells were left untreated, treated with TPA or U0126 or pre-treated with 5 μM SB 203580 for 1 hour and then left untreated or treated with TPA or U0126 for the times indicated. Immunoblots of total lysates were performed using specific antibodies recognising p21 WAF1 and α-tubulin (A) and pRb (B). Densitometric analysis of bands provided quantification expressed as the ratio of amount of p21 WAF1 versus α-tubulin amount. (C) Northern blot analysis of p21 WAF1 , myogenin and MyoD expression in RD cells left untreated or treated with U0126 in the absence or in the presence of SB 203580 for the times indicated. The GAPDH mRNA levels are shown. The data shown are representative of three independent experiments for A and B and two for C.

Article Snippet: The p21 WAF1 , MyoD and myogenin probes was obtained from the plasmid described below, while cyclin D1 probe was kindly provided by Dr. A. Arnold [ ] and GAPDH vector was provided by ATCC.

Techniques: Inhibition, Expressing, Western Blot, Northern Blot

Journal: Molecular Cancer

Article Title: p21 WAF1 expression induced by MEK/ERK pathway activation or inhibition correlates with growth arrest, myogenic differentiation and onco-phenotype reversal in rhabdomyosarcoma cells

doi: 10.1186/1476-4598-4-41

Figure Lengend Snippet: p21 WAF1 expression induced by U0126 is dependent on MyoD and/or myogenin. (A) Immunoblots of total lysates from RD cells transiently transfected with control-, MyoD- or myogenin-siRNA, or with a combination of MyoD- and myogenin-siRNA (MyoD+myogenin), and then left untreated (-) or treated with U0126 (+) for 2 days. (B) Immunoblots of total lysates from Puromycin-selected polyclonal population from RD cells transfected with the empty vector (CMV), MyoD- or myogenin-expressing vector. As a control, cells were left untransfected in the absence (C) or in the presence of TPA (TPA). Immunoblots were performed using specific antibodies capable of recognising MyoD, myogenin, p21 WAF1 and α-tubulin. Densitometric analysis of bands provided quantification of MyoD and myogenin levels expressed as a fold increase over the control value (CMV) arbitrarily set at 1. (C) Luciferase assay of lysates from RD cells co-transfected with the empty vector (CMV), myogenin- or MyoD-expressing vector and the plasmid carrying p21 WAF1 promoter (DM-Luc). Data show mean values ± s.e.m. of triplicates of a representative experiment. Similar results were obtained in two experiments.

Article Snippet: The p21 WAF1 , MyoD and myogenin probes was obtained from the plasmid described below, while cyclin D1 probe was kindly provided by Dr. A. Arnold [ ] and GAPDH vector was provided by ATCC.

Techniques: Expressing, Western Blot, Transfection, Plasmid Preparation, Luciferase

Journal: Molecular Cancer

Article Title: p21 WAF1 expression induced by MEK/ERK pathway activation or inhibition correlates with growth arrest, myogenic differentiation and onco-phenotype reversal in rhabdomyosarcoma cells

doi: 10.1186/1476-4598-4-41

Figure Lengend Snippet: Induction of p21 WAF1 expression and growth arrest in RH30 cells treated with TPA and U0126. (A) RH30 cells treated with 10 -7 M TPA (T) or 10 μM U0126 (U) for the times indicated. Whole cell lysates were analysed by immunoblotting with a specific antibody for p21 WAF1 , phosphorylated ERK1/2 and α-tubulin expression. (B) Growth graph of RH30 and RD cells after 4 days of TPA (T) or U0126 (U) treatments. (C) Immunoblot of total lysates using specific antibody capable of recognising the sarcomeric myosin heavy chain (MHC). Similar results were obtained in two experiments.

Article Snippet: The p21 WAF1 , MyoD and myogenin probes was obtained from the plasmid described below, while cyclin D1 probe was kindly provided by Dr. A. Arnold [ ] and GAPDH vector was provided by ATCC.

Techniques: Expressing, Western Blot

Journal: Molecular Cancer

Article Title: p21 WAF1 expression induced by MEK/ERK pathway activation or inhibition correlates with growth arrest, myogenic differentiation and onco-phenotype reversal in rhabdomyosarcoma cells

doi: 10.1186/1476-4598-4-41

Figure Lengend Snippet: Enforced p21 WAF1 expression induces growth arrest in RD cells. (A) Immunoblotting using specific antibody capable of recognising the p21 WAF1 of total lysates from RD cells transfected with CB6/p21 or the empty vector (CB6), cultured without (-) or with 120 μM ZnCl 2 (+) for 3 days before (Transient transfection) and after (Stable transfection) neomycin-selection. (B) Growth graph of same neomycin-selected RD cells cultured as in A. As a control, untransfected RD cells were left untreated (C) or treated with TPA or U0126. (C) Cell cycle distribution of RD cells transfected with empty vector PEGFP ( left ) or with PEGFP/p21 ( right ). Cell cycle distribution of GFP-expressing cells was evaluated by flow cytometry. The data shown in the insert tables are the mean ± s.e.m. of triplicates of a representative experiment. Similar results were obtained in two experiments.

Article Snippet: The p21 WAF1 , MyoD and myogenin probes was obtained from the plasmid described below, while cyclin D1 probe was kindly provided by Dr. A. Arnold [ ] and GAPDH vector was provided by ATCC.

Techniques: Expressing, Western Blot, Transfection, Plasmid Preparation, Cell Culture, Stable Transfection, Selection, Flow Cytometry

Journal: Molecular Cancer

Article Title: p21 WAF1 expression induced by MEK/ERK pathway activation or inhibition correlates with growth arrest, myogenic differentiation and onco-phenotype reversal in rhabdomyosarcoma cells

doi: 10.1186/1476-4598-4-41

Figure Lengend Snippet: Enforced p21 WAF1 expression induces reversion of anchorage independent growth. Neomycin-selected RD cells transfected with CB6/p21 or the empty vector (CB6) were suspended in 0.33% Difco agar in the absence (CB6, CB6/p21) or presence (CB6 Zn 2+ , CB6/p21 Zn 2+ ) of 120 μM ZnCl 2 and overlaid on an 0.5% agar layer. Colonies were photographed after 14 days. Similar results were obtained in two experiments.

Article Snippet: The p21 WAF1 , MyoD and myogenin probes was obtained from the plasmid described below, while cyclin D1 probe was kindly provided by Dr. A. Arnold [ ] and GAPDH vector was provided by ATCC.

Techniques: Expressing, Transfection, Plasmid Preparation