Journal: Cancer research

Article Title: Novel mechanism of MDA-7/IL-24 cancer-specific apoptosis through SARI induction

doi: 10.1158/0008-5472.CAN-13-1062

Figure Lengend Snippet: The indicated cells were infected as indicated with Ad.vec or Ad.mda-7 (or Ad.p53) at 20 pfu/cell (HeLa, LNCaP) or 100 pfu/cell (other cells) for 24 h. For Quantitative mRNA detection, total RNA was purified, blotted onto nylon membranes and Northern blotting was done with a SARI cDNA. A) Im-PHFA, malignant glioma and neuroglioma cells. B) P69, prostate cancer and HeLa cells. C) FM516 and melanoma cells. GAPDH mRNA was used as loading control.

Article Snippet: Cell lines and stable clones Normal SV-40-immortalized human prostate epithelial cells (P69) were obtained from Dr. Ware, VCU, and DU-145, PC3 and LNCaP human prostate carcinoma cells were obtained from American Type Culture Collection (ATCC) and cultured as described ( 25 ).

Techniques: Infection, Purification, Northern Blot, Control

Journal: Cancer research

Article Title: Novel mechanism of MDA-7/IL-24 cancer-specific apoptosis through SARI induction

doi: 10.1158/0008-5472.CAN-13-1062

Figure Lengend Snippet: The indicated cell lines were infected with Ad.vec or Ad.mda-7 at 100 pfu/cell for the indicated time after which cell lysates were collected. For detecting protein expression, 50 μg of total protein was run on SDS–PAGE, transferred onto a nitrocellulose membrane and stained with the indicated antibodies, and protein expression was determined by Western blotting in A) Im-PHFA, malignant glioma and neuroglioma, B) P69 and prostate cancer cells, and C) FM516 and melanomas. D) Cells were infected with Ad.vec or Ad.mda-7 at 100 pfu/cell for the indicated time after which cell proliferation/viability was assayed by MTT assay. All experiments were performed at least three times, and data represent mean ± S.D. * p<0.05 cell viability is significantly less than cells infected for 0 h (one way ANOVA with Newman Keuls).

Article Snippet: Cell lines and stable clones Normal SV-40-immortalized human prostate epithelial cells (P69) were obtained from Dr. Ware, VCU, and DU-145, PC3 and LNCaP human prostate carcinoma cells were obtained from American Type Culture Collection (ATCC) and cultured as described ( 25 ).

Techniques: Infection, Expressing, SDS Page, Membrane, Staining, Western Blot, MTT Assay

Journal: Cancer research

Article Title: Novel mechanism of MDA-7/IL-24 cancer-specific apoptosis through SARI induction

doi: 10.1158/0008-5472.CAN-13-1062

Figure Lengend Snippet: A) SARI full length promoter/pGL3-Luc or SARI deletion mutant promoter/PGL3-Luc constructs were transfected into HeLa cells. Twenty-four h later the cells were treated with IFN-β (1000 U/ml) or infected with Ad.vec (100 pfu/cell) or Ad.mda-7 (100 pfu/cell) for 24 h after which cell lysates were collected for luciferase assays. B) i): HeLa cells were infected with Ad.vec or Ad.SARI at 10 pfu/cell and nuclei were prepared from the treated cells. The isolated nuclei were used to label preinitiated RNA transcription with [α-32P] UTP in vitro, and the purified RNA was hybridized to a dot blot carrying an equivalent amount of a SARI cDNA. The transcription rate of GAPDH served as control. ii) HeLa cells were treated as described in panel A and total RNA was purified, blotted onto a nylon membrane, and Northern blotting was done with a SARI, mda-7/IL-24 or GAPDH cDNA. C) 3′-UTR region of SARI mRNA was cloned into PGL3-basic-Luc vector (Luc-3′UTR) and three independent clones (along with control vector) were transfected into HeLa cells for 24 h followed by infection with Ad.vec or Ad.mda-7 (100 pfu/cell for 24 h) and cell lysates were collected for luciferase assays. The data is presented as the ratio of Ad.mda-7 to Ad.vec treated cells. All experiments were performed at least three times, and data represent mean ± S.D. * p<0.05 RLU is significantly higher than PGL-3 Luc control cells. D) P69 cells were infected with Ad.vec or Ad.mda-7 (20 pfu/cell for 24 h) after which Actinomycin D (0.5 μg/ml) was added for the indicated times. Total RNA was collected to perform Northern blotting using a SARI cDNA.

Article Snippet: Cell lines and stable clones Normal SV-40-immortalized human prostate epithelial cells (P69) were obtained from Dr. Ware, VCU, and DU-145, PC3 and LNCaP human prostate carcinoma cells were obtained from American Type Culture Collection (ATCC) and cultured as described ( 25 ).

Techniques: Mutagenesis, Construct, Transfection, Infection, Luciferase, Isolation, In Vitro, Purification, Dot Blot, Control, Membrane, Northern Blot, Clone Assay, Plasmid Preparation

Journal: Cell Reports Medicine

Article Title: Unraveling AURKB as a potential therapeutic target in pulmonary hypertension using integrated transcriptomic analysis and pre-clinical studies

doi: 10.1016/j.xcrm.2025.101964

Figure Lengend Snippet: Therapeutic effects of barasertib in male rats exposed to Sugen/hypoxia (A) Study design using the Sugen/hypoxia (Su/Hx) rat model. (B) Pulmonary artery acceleration time (PAAT), right ventricular fractional area change (RVFAC), tricuspid annular plane systolic excursion (TAPSE), S wave, stroke volume (SV), and cardiac output (CO) determined by echocardiography at the end of the protocol in control, Su/Hx+Veh, and Su/Hx+barasertib male rats ( n = 4–9/group). (C) Effect of AURKB inhibition on right ventricular systolic pressure (RVSP) and mean pulmonary artery pressure (mPAP), as assessed by right heart catheterization at the end of the protocol ( n = 4–9/group). (D) Representative images of distal PAs stained with Elastica van Gieson (EVG) and quantification of vascular remodeling in control, Su/Hx+Veh, and Su/Hx+barasertib rats ( n = 4–9/group). (E) Representative images of distal PAs labeled with proliferating cell nuclear antigen (PCNA, proliferative marker, red), p16, or p21 ( n = 4–9/group). PASMCs were labeled with alpha smooth muscle actin (αSMA, green). The quantifications of the percentage of PASMCs positive for PCNA, p16, or p21 are shown. Scale bars, 20 μm. Scatter dot plots show individual values and mean ± SEM. Assessment of the normality of the data was performed using Shapiro-Wilk test. Statistical analyses were performed using one-way ANOVA or Kruskal-Wallis’s test followed by Dunnett’s post hoc test; ∗ p < 0.05; ∗∗ p < 0.01, and ∗∗∗ p < 0.001. See also Figures S8 and .

Article Snippet: Smooth muscle cell basal medium , Cell Applications , Cat# 310-500.

Techniques: Control, Inhibition, Staining, Labeling, Marker

Journal: Cell Reports Medicine

Article Title: Unraveling AURKB as a potential therapeutic target in pulmonary hypertension using integrated transcriptomic analysis and pre-clinical studies

doi: 10.1016/j.xcrm.2025.101964

Figure Lengend Snippet: Barasertib reduces vascular remodeling in human precision-cut lung slices (A) Experimental setup for precision-cut lung slices (PCLSs) from control, patients with PAH, and patients with idiopathic pulmonary fibrosis. (B) Representative images of distal PAs stained with Elastica van Gieson (EVG) or labeled with proliferating cell nuclear antigen (PCNA) or p21 in PCLSs prepared from control patients ( n = 5) after exposure or not to a growth factor cocktail (GF, FGF2 + PDGF-BB + ET1) in presence or not to barasertib for 10 days. PASMCs were labeled with alpha smooth muscle actin (αSMA, green). The quantification of vascular remodeling and PASMCs positive for PCNA or p21 is shown. (C) Representative images of distal PAs stained with EVG or labeled with PCNA or p21 in PCLSs from patients with PAH ( n = 6). (D) Representative images of distal PAs stained with EVG or labeled with PCNA or p21 in PCLSs from patients with IPF complicated with pulmonary hypertension (PH) ( n = 2). For each experiment, the quantifications of vascular remodeling and PCNA- or p21-positive PASMCs (average of 40–45 arteries per patient) are shown. Scale bars, 25 μm. Values are represented as means ± SEM. Assessment of the normality of the data was performed using Shapiro-Wilk test. Statistical analyses were performed using repeated measures one-way ANOVA test followed by Dunnett’s post hoc test or paired Student’s t test; ∗ p < 0.05; ∗∗ p < 0.01, and ∗∗∗ p < 0.001.

Article Snippet: Smooth muscle cell basal medium , Cell Applications , Cat# 310-500.

Techniques: Control, Staining, Labeling

Journal: Cell Reports Medicine

Article Title: Unraveling AURKB as a potential therapeutic target in pulmonary hypertension using integrated transcriptomic analysis and pre-clinical studies

doi: 10.1016/j.xcrm.2025.101964

Figure Lengend Snippet:

Article Snippet: Smooth muscle cell basal medium , Cell Applications , Cat# 310-500.

Techniques: Virus, Plasmid Preparation, Recombinant, Negative Control, Protease Inhibitor, Staining, EdU Assay, TUNEL Assay, Western Blot, SYBR Green Assay, Chromatin Immunoprecipitation, Magnetic Beads, RNA Sequencing Assay, Expressing, Software