Journal: FEBS Open Bio

Article Title: Reduced MAGI3 level by HPV18E6 contributes to Wnt/β‐catenin signaling activation and cervical cancer progression

doi: 10.1002/2211-5463.13298

Figure Lengend Snippet: MAGI3 downregulates β‐catenin protein level and inhibits Wnt/β‐catenin‐mediated signaling in CC cells. (A) MAGI3 overexpression retarded Wnt signaling activation via reducing β‐catenin protein level. HeLa‐MAGI3 cells were treated in the presence (lanes 3 and 4) or absence (lanes 1 and 2) of Wnt/β‐catenin activator CHIR‐99021 (0.2 µmol·L −1 ) for 4 h. The expression levels of β‐catenin and TCF1 were analyzed by western blotting. (B) MAGI3 knockdown promoted Wnt signaling activation via increasing β‐catenin level. C‐33A cells were transfected with MAGI3 siRNAs or scrambles for 24 h and then treated with Wnt/β‐catenin inhibitor IWR‐1‐endo (100 µmol·L −1 ; lanes 3 and 4) for 24h, and cell lysates were analyzed by western blotting. The gray value of protein bands of western blots has been quantified by imagej . All experiments were repeated three times ( n = 3).

Article Snippet: CHIR‐99021, a Wnt/β‐catenin activator, was purchased from TargetMol (Boston, MA, USA); IWR‐1‐endo, a Wnt/β‐catenin inhibitor, was purchased from Selleck (Houston, TX, USA). pcDNA3‐V5/His‐MAGI3 and GFP/GFP‐MAGI3 plasmids were kindly provided by R. Hall (Emory University, GA).

Techniques: Over Expression, Activation Assay, Expressing, Western Blot, Transfection

Journal: FEBS Open Bio

Article Title: Reduced MAGI3 level by HPV18E6 contributes to Wnt/β‐catenin signaling activation and cervical cancer progression

doi: 10.1002/2211-5463.13298

Figure Lengend Snippet: Downregulation of MAGI3 by HPV18 E6 rather than HPV16 E6 enhances cell migration and invasion via activation of β‐catenin signaling. (A, B) knockdown of HPV18 E6 inhibited the migration and invasion of HeLa cells via increasing MAGI3 and reducing β‐catenin level. HPV18 E6 was knocked down by HPV18 E6 siRNAs, or codepleted with MAGI3 siRNAs in the presence or absence of β‐catenin knockdown in HeLa cells. (C, D) Overexpression of HPV18 E6 promoted the migration and invasion of C‐33A cells via reducing MAGI3 and increasing β‐catenin level. HPV18 E6 was overexpressed alone or co‐overexpressed with MAGI3 in the presence or absence of Wnt/β‐catenin activator CHIR‐99021 in C‐33A cells. (E) HPV18 E6 rather than HPV16 E6 promoted CC cell migration and invasion through activating Wnt/β‐catenin signaling. HPV18 E6 and HPV16 E6 were overexpressed in C‐33A cells, or co‐treated with Wnt/β‐catenin inhibitor IWR‐1‐endo or vehicle. Cell lysates, migration and invasion were conducted as described in Fig. . The expression level of protein was quantitatively expressed by relative gray value, all experiments were repeated three times ( t‐ test; * P < 0.05, ** P < 0.01, and *** P < 0.001, values represent mean ± SD, n = 3). Scale bar, 100 μm.

Article Snippet: CHIR‐99021, a Wnt/β‐catenin activator, was purchased from TargetMol (Boston, MA, USA); IWR‐1‐endo, a Wnt/β‐catenin inhibitor, was purchased from Selleck (Houston, TX, USA). pcDNA3‐V5/His‐MAGI3 and GFP/GFP‐MAGI3 plasmids were kindly provided by R. Hall (Emory University, GA).

Techniques: Migration, Activation Assay, Over Expression, Expressing