Quantitative Analysis of RhoA Dynamics

Human breast cancer cell line MDA-MB-231 was purchased from ATCC. RhoA antibody (sc-418) and donkey anti-mouse IgG-FITC (sc-2099) were purchased from Santa Cruz. Duramycin, biotin and phalloidin tetramethylrhodamine (Cat#P1951) were from Sigma; biotinylated Duramycin was synthesised as described earlier (8 (link)); avidin-Texas red conjugate (A820) and avidin-FITC (Cat#43-4411) were from Life Technologies; Zeba desalt spin column (Cat#89890) was from Thermo Scientific. GFP-tagged wild-type RhoA (GFP-RhoA), constitutively active RhoA (GFP-CA-RhoA), wild-type RhoA with positive charges in the PBR mutated to Q (GFP-RhoA-PBRQ) and constitutively active RhoA with positive charges in the PBR mutated to Q (GFP-CA-RhoA-PBRQ) were prepared and used as previously described (9 (link)). Lact-C2-GFP (Cat#22852), 2PH-PLCdelta-GFP (Cat#35142) and PM-GFP (Cat#21213) were purchased from Addgene. LifeAct–TagGFP2 (Cat#60101) was purchased from Ibidi.

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Hou S., Grillo D., Williams C.L., Wasserstrom J.A., Szleifer I, & Zhao M. (2014). Membrane phospholipid redistribution in cancer micro-particles and implications in the recruitment of cationic protein factors. Journal of Extracellular Vesicles, 3, 10.3402/jev.v3.22653.

Publication 2014

MDA-MB-231 donkey anti-mouse IgG-FITC (sc-2099) Duramycin biotin phalloidin tetramethylrhodamine avidin-FITC Zeba desalt spin column Lact-C2-GFP PM-GFP LifeAct–TagGFP2

Anti igg Antibody Avidin Biotin Breast cancer cell line Donkey Duramycin Fitc Human Mouse Phalloidin Rhoa Tetramethylrhodamine

Corresponding Organization : Northwestern University

Other organizations : Medical College of Wisconsin

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Variable analysis

independent variables

GFP-tagged wild-type RhoA (GFP-RhoA)
Constitutively active RhoA (GFP-CA-RhoA)
Wild-type RhoA with positive charges in the PBR mutated to Q (GFP-RhoA-PBRQ)
Constitutively active RhoA with positive charges in the PBR mutated to Q (GFP-CA-RhoA-PBRQ)

dependent variables

Not explicitly mentioned

control variables

Human breast cancer cell line MDA-MB-231 from ATCC
RhoA antibody (sc-418) from Santa Cruz
Donkey anti-mouse IgG-FITC (sc-2099) from Santa Cruz
Duramycin, biotin and phalloidin tetramethylrhodamine (Cat#P1951) from Sigma
Biotinylated duramycin synthesised as described earlier (8 (link))
Avidin-Texas red conjugate (A820) and avidin-FITC (Cat#43-4411) from Life Technologies
Zeba desalt spin column (Cat#89890) from Thermo Scientific
Lact-C2-GFP (Cat#22852), 2PH-PLCdelta-GFP (Cat#35142) and PM-GFP (Cat#21213) from Addgene
LifeAct–TagGFP2 (Cat#60101) from Ibidi

controls

Positive controls: Not explicitly mentioned
Negative controls: Not explicitly mentioned

Annotations

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