Western Blot and IHC Analysis of Metastatic Markers

Immunoblotting and IHC analysis were conducted with standard procedures, as previously described 15 (link), 16 (link). Blotting membranes were stripped and reprobed with anti-β-Actin antibody as a loading control. The degree of immunostaining of formalin-fixed, paraffin-embedded sections was reviewed and scored independently by two pathologists based on both the proportion of positively stained tumor cells and the intensity of the staining. The following antibodies were used for immunoblotting or IHC analysis: ARHGAP5 (#2562, WB, 1:1000), Vimentin (#5741; WB, 1:1000; IHC, 1:200), N-cadherin (#13116; WB, 1:1000; IHC, 1:100), E-cadherin (#14472; WB, 1:1000; IHC, 1:100), RhoA (#2171; WB, 1:1000), CREB1 (#9197; WB, 1:1000; IHC, 1:6000), β-Actin (#3700; WB, 1:3000) (all from Cell Signaling, Beverly, USA) and ARHGAP5 (#ab199160; IHC, 1:100) (Abcam, Cambridge, MA, USA).

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Tian T., Chen Z.H., Zheng Z., Liu Y., Zhao Q., Liu Y., Qiu H., Long Q., Chen M., Li L., Xie F., Luo G., Wu X, & Deng W. (2020). Investigation of the role and mechanism of ARHGAP5-mediated colorectal cancer metastasis. Theranostics, 10(13), 5998-6010.

Publication 2020

ARHGAP5 β-Actin

Actin Anti antibody Antibodies E cadherin Embedded paraffin Formalin Membranes N cadherin Pathologists Rhoa Tumor Vimentin

Corresponding Organization :

Other organizations : Sun Yat-sen University Cancer Center, Sun Yat-sen University, Third Affiliated Hospital of Sun Yat-sen University

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Variable analysis

independent variables

Immunoblotting and IHC analysis were conducted with standard procedures, as previously described 15 (link), 16 (link).
The following antibodies were used for immunoblotting or IHC analysis: ARHGAP5 (#2562, WB, 1:1000), Vimentin (#5741; WB, 1:1000; IHC, 1:200), N-cadherin (#13116; WB, 1:1000; IHC, 1:100), E-cadherin (#14472; WB, 1:1000; IHC, 1:100), RhoA (#2171; WB, 1:1000), CREB1 (#9197; WB, 1:1000; IHC, 1:6000), β-Actin (#3700; WB, 1:3000) (all from Cell Signaling, Beverly, USA) and ARHGAP5 (#ab199160; IHC, 1:100) (Abcam, Cambridge, MA, USA).

dependent variables

The degree of immunostaining of formalin-fixed, paraffin-embedded sections was reviewed and scored independently by two pathologists based on both the proportion of positively stained tumor cells and the intensity of the staining.

control variables

Blotting membranes were stripped and reprobed with anti-β-Actin antibody as a loading control.

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