Immunofluorescence and Cell Assays Protocol

The following antibodies were used in this study: SHROOM2 (SAB2700199), Vinculin (V9131) and Phalloidin–Tetramethylrhodamine B isothiocyanate (P1951) were from Sigma; E-Cadherin (24E10), N-Cadherin (13116), MLC2 (3672), p-MLC2 (T18/S19) (3674), HA (3724) were from Cell Signaling Technology; ROCK1 (ab58305), CK5 (ab52635), CK10 (ab76318), Desmoglein1 (ab12077) and RAC1 (ab155938) were from abcam; p-MYPT (ABS45) was from Millipore and Fibronectin (610077) was from BD biosciences. Horseradish peroxidase-conjugated goat anti-mouse/rabbit secondary antibodies were purchased from Promega.
Immunofluorescence microscopy, cell proliferation, colony formation, soft agar assays were performed as described previously^{8 (link),42 (link)}.

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Yuan J., Chen L., Xiao J., Qi X.K., Zhang J., Li X., Wang Z., Lian Y.F., Xiang T., Zhang Y., Chen M.Y., Bei J.X., Zeng Y.X, & Feng L. (2019). SHROOM2 inhibits tumor metastasis through RhoA–ROCK pathway-dependent and -independent mechanisms in nasopharyngeal carcinoma. Cell Death & Disease, 10(2), 58.

Publication 2019

Phalloidin–Tetramethylrhodamine B isothiocyanate (P1951) Fibronectin (610077) Horseradish peroxidase-conjugated goat anti-mouse/rabbit secondary antibodies

Agar Anti antibodies Antibodies Assays Cell proliferation E cadherin Fibronectin Goat Horseradish peroxidase Immunofluorescence microscopy Mouse N cadherin Phalloidin Promega Rabbit Rock1 Tetramethylrhodamine isothiocyanate Vinculin

Corresponding Organization :

Other organizations : Sun Yat-sen University, Sun Yat-sen University Cancer Center, Westlake University

Top 5 similar protocols

Variable analysis

independent variables

SHROOM2 (SAB2700199)
Vinculin (V9131)
Phalloidin–Tetramethylrhodamine B isothiocyanate (P1951)
E-Cadherin (24E10)
N-Cadherin (13116)
MLC2 (3672)
P-MLC2 (T18/S19) (3674)
HA (3724)
ROCK1 (ab58305)
CK5 (ab52635)
CK10 (ab76318)
Desmoglein1 (ab12077)
RAC1 (ab155938)
P-MYPT (ABS45)
Fibronectin (610077)

dependent variables

Immunofluorescence microscopy
Cell proliferation
Colony formation
Soft agar assays

control variables

Horseradish peroxidase-conjugated goat anti-mouse/rabbit secondary antibodies

Annotations

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