Journal:

Article Title: Positional enhancer-blocking activity of the chicken ?-globin insulator in transiently transfected cells

doi:

Figure Lengend Snippet: (A) Transient activity in 6C2 cells of the different fragments of the chicken β-globin insulator. An enhancer-blocking assay was performed to test the ability of the 1.2-kb DNA fragment (constructs 2d and 2e), the core (250-bp; constructs 2b and 2c), and (II/III-Ins)Q to block the action of an enhancer (construct 2a). All constructs have a SV40 enhancer (open box) and a SV40 promoter (striped box) driving a luciferase reporter gene (construct 2f). Test fragments are located between the enhancer and promoter (constructs 2a–2e). The distance between the enhancer and its promoter is indicated. Error bars represent the standard error of the means (SEM) for seven experiments; (+) and (−) symbols show the 5′ to 3′ and 3′ to 5′ orientation of the fragments, respectively. (B) Transient activity of circular plasmids in 6C2 cells of the (II/III-Ins)Q element with various enhancer–promoter constructions in the pGL2 series of luciferase vectors. The reduction in reporter activity (7.4-fold) was caused by the insulator (construct 2h); the reduction in enhancer activity (2.4-fold) relative to the SV40 promoter alone (compare construct 2g with 2i). Data are the average of five independent experiments; error bars show the SEM here and in all figures.

Article Snippet: The chicken (II/III-Ins)Q fragment (Fig. ) was liberated from pNI-II/IIIQ by digestion with Bam HI and Eco RI and cloned in the Xho I site of the pGL2-control vector (Promega) by using Xho I-linkers (New England Biolabs).

Techniques: Activity Assay, Blocking Assay, Construct, Luciferase

Journal: Cell Proliferation

Article Title: p62 functions as an oncogene in colorectal cancer through inhibiting apoptosis and promoting cell proliferation by interacting with the vitamin D receptor

doi: 10.1111/cpr.12585

Figure Lengend Snippet: p62 shows tumour‐promoting ability in CRC cells. A, Western blotting of p62 in 6 human CRC cell lines and one normal human intestinal epithelial cell line. B, HCT116 was stably transduced with a p62‐overexpression lentiviral vector (p62‐OE), and SW480 was stably transduced with a p62‐knockdown lentiviral vector (p62‐KD). Accordingly, control groups were transduced with the corresponding lentiviral carrying an empty vector (EV). Overexpression or silencing of p62 was confirmed by RT‐qPCR (left panel) and Western blotting (right panel). C, Overexpression of p62 significantly promoted cell viability by CCK‐8 assay (left panel) compared to that of the control groups in HCT116. Knockdown of p62 significantly reduced cell viability by CCK‐8 assay (right panel) compared to that of the controls in SW480. D, Overexpression of p62 significantly increased colony numbers (left panel) compared to those of the control groups in HCT116. Knockdown of p62 significantly decreased colony numbers (right panel) compared to those of the controls in SW480. E, Apoptosis was analysed by flow cytometry. Knockdown of p62 in SW480 cells induced early apoptosis compared to the control cells. F, Expression of the cleaved form and the total level of caspase‐7 and PARP1 were detected by Western blotting. Bars represent the standard error of the mean ± SD from three independent experiments. *Represents Student's t test * P < 0.05, ** P < 0.01 and *** P < 0.001.

Article Snippet: The hU6‐MCS‐CMV‐EGFP knockdown lentiviral vector for SQSTM1 , the Ubi‐MCS‐3FLAG‐SV40‐puromycin overexpressing lentiviral vector for SQSTM1 and the control lentiviral vector were obtained from Genechem Co., Ltd. (Shanghai, China).

Techniques: Western Blot, Stable Transfection, Transduction, Over Expression, Plasmid Preparation, Quantitative RT-PCR, CCK-8 Assay, Flow Cytometry, Expressing