Journal: Molecular cell

Article Title: MYC assembles and stimulates topoisomerases 1 and 2 in a “topoisome”

doi: 10.1016/j.molcel.2021.11.016

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Mouse monoclonal c-Myc Antibody (9E10) , Santa Cruz Biotechnology , Cat# sc-40 AC; RRID: AB_2857941.

Techniques: Recombinant, Control, Affinity Purification, Virus, Cell Culture, Protease Inhibitor, Magnetic Beads, Transfection, Plasmid Preparation, Titration, Staining, Thin Layer Chromatography, SYBR Green Assay, In Situ, Purification, Sample Prep, Sequencing, RNA Sequencing, Cloning, CRISPR, Inverse PCR, Expressing, Software

Journal:

Article Title: Myc Downregulation by Transforming Growth Factor ? Required for Activation of the p15 Ink4b G 1 Arrest Pathway

doi:

Figure Lengend Snippet: Generation of Mv1Lu cell lines with inducible c-Myc expression. (A) Clonal tet-Myc cell lines (TM1 to TM3) were maintained in medium containing 1 μg of tetracycline per ml and then grown in the absence of tetracycline for 18 h before being harvested and analyzed by anti-human c-Myc Western immunoblotting. Parental Mv1Lu cells were also analyzed in the presence and absence of tetracycline with the anti-human c-Myc antibody. (B) Comparison of the levels of induced human c-Myc expressed in tet-Myc clones with other human cell lines by anti-human c-Myc Western immunoblotting. The same amount of cell lysate protein was loaded in each lane. (C) The human c-Myc protein levels visualized in panel B were quantitated by densitometry of the signal and plotted as relative levels of c-Myc protein in arbitrary units.

Article Snippet: Antibodies against human c-Myc (sc-40, 9E10), cyclin D1 (sc-753, sc-718), cdk6 (sc-177, C21), cyclin D2 (sc-181), or human p15 (sc-612) were purchased from Santa Cruz Biotechnology.

Techniques: Expressing, Western Blot, Clone Assay

Journal:

Article Title: Myc Downregulation by Transforming Growth Factor ? Required for Activation of the p15 Ink4b G 1 Arrest Pathway

doi:

Figure Lengend Snippet: Effect of exogenous c-Myc expression on G1 cdk components. TM2 cells were induced to express exogenous Myc by the removal of tetracycline and harvested at the indicated times. Immunoblots were probed for exogenous human c-Myc and endogenous mink cdk2, p27, cdk4, cdk6, cyclin D1, and cyclin D2 proteins.

Article Snippet: Antibodies against human c-Myc (sc-40, 9E10), cyclin D1 (sc-753, sc-718), cdk6 (sc-177, C21), cyclin D2 (sc-181), or human p15 (sc-612) were purchased from Santa Cruz Biotechnology.

Techniques: Expressing, Western Blot

Journal:

Article Title: Myc Downregulation by Transforming Growth Factor ? Required for Activation of the p15 Ink4b G 1 Arrest Pathway

doi:

Figure Lengend Snippet: Induction of p15Ink4b in tet-Myc clones. (A and B) Three tet-Myc clones (TM1 to TM3) were induced to express c-Myc by the absence of tetracycline for 18 h and then treated with 200 pM TGF-β. At 6 h after TGF-β addition, the cells were harvested and their RNA was analyzed by Northern blotting for p15Ink4b expression. A representative blot with the TM1 cell lines is shown (A). The same blot was probed for GAPDH expression as a loading control. Quantitation of such p15 blots for all three TM clones was carried out and is shown as the fold induction of this RNA by addition of TGF-β, normalized to GAPDH values (B). (C) p15Ink4b induction in parental Mv1Lu cells and TM1 cells after 20 h without tetracycline followed by TGF-β addition for the indicated times. The same Northern blot was probed for PAI-1 as another representative TGF-β-responsive gene and GAPDH as a loading control. (D) TM1 cells were transfected with the p15p113-Luc reporter. Transfections were carried out without or with TGFβ in the absence or presence of tetracycline for 24 h prior to analysis of luciferase activity. Results are means ± standard deviations of triplicate transfections. HaCaT cells were transfected with p15 p113-Luc reporter, without or with a human c-Myc expression vector, and analyzed as described above.

Article Snippet: Antibodies against human c-Myc (sc-40, 9E10), cyclin D1 (sc-753, sc-718), cdk6 (sc-177, C21), cyclin D2 (sc-181), or human p15 (sc-612) were purchased from Santa Cruz Biotechnology.

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

Journal: Journal of Biological Chemistry

Article Title: Two Functionally Divergent UDP-Gal Nucleotide Sugar Transporters Participate in Phosphoglycan Synthesis in Leishmania major

doi: 10.1074/jbc.m610869200

Figure Lengend Snippet: FIGURE 5. LPG5A and LPG5B rescue M. amurensis reactivity in the UDP- Gal transporter-deficient Lec8 mutant. Part A, fluorescence microscopy. A–F display the fluorescent images of cells stained with fluorescein iso- thiocyanate-labeled M. amurensis lectin, and G–L display the phase-con- trast images. A and G, WT CHO cells. B and H, Lec8/hUGT1. C and I, Lec8/ empty; D and J, Lec8/LPG5A; E and K, Lec8/LPG5B; F and L, Lec8/HUT1L. Part B, flow cytometry. Stably transfected Lec8 CHO cells were fixed and incubated with fluorescein isothiocyanate-labeled M. amurensis lectin. In A–E, WT or transfected Lec8 cells (solid black lines) are shown compared with Lec8 transfected with empty vector (Lec8; dotted line). These data are representative of at least two independent experiments.

Article Snippet: Heterologous Expression—WT Chinese hamster ovary (CHO) and Lec8 cells (40, 41) were from ATCC (Manassas, VA) and maintained in culture following ATCC recommendations.

Techniques: Mutagenesis, Fluorescence, Microscopy, Staining, Labeling, Flow Cytometry, Stable Transfection, Transfection, Incubation, Plasmid Preparation