Gene Regulation Techniques in Cell Biology

siRNA against WASF3 and the pLKO lentiviral vectors containing an shRNA against SHOX2 or STAT3 were purchased from Open Biosystems (Huntsville, AL). Coding sequences of human HA-tagged WASF3 and FLAG-tagged SHOX2 were respectively cloned into pCDH-CMV-MCS-EF1 lentivector (System Biosciences, Mountain View, CA) as described previously [8 (link), 13 (link)]. The pGL-WASF3 promoter constructs (-350/+494, -650/+494, -1250/+494) were generated as described previously [16 (link)]. The following primary antibodies were used in Western blot assays: anti-WASF3, anti-STAT3, anti-pSTAT3 (Tyr 705), and anti-E-cadherin antibodies (Cell Signaling Technology, MA); anti-HA, anti-FLAG, and anti-β-Actin antibodies (Sigma-Aldrich, MO); and anti-SHOX2 antibody (Abcam, MA). The STAT3 inhibitor S3I-201 was obtained from Selleckchem (Houston, TX).

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Teng Y., Loveless R., Benson E.M., Sun L., Shull A.Y, & Shay C. (2021). SHOX2 cooperates with STAT3 to promote breast cancer metastasis through the transcriptional activation of WASF3. Journal of Experimental & Clinical Cancer Research : CR, 40, 274.

Publication 2021

pLKO lentiviral vectors anti-WASF3 anti-STAT3 anti-pSTAT3 (Tyr 705), anti-E-cadherin antibodies anti-SHOX2 antibody

Actin Anti antibodies Anti antibody Antibodies Cadherin Coding sequences Human S3i 201 Shrna Sirna Stat3 Vectors Wasf3 Western blot assays

Corresponding Organization : Emory University

Other organizations : Augusta University Health, Presbyterian College, Children's Center

Top 5 similar protocols

Variable analysis

independent variables

SiRNA against WASF3
PLKO lentiviral vectors containing an shRNA against SHOX2
PLKO lentiviral vectors containing an shRNA against STAT3
HA-tagged WASF3
FLAG-tagged SHOX2
PGL-WASF3 promoter constructs (-350/+494, -650/+494, -1250/+494)
STAT3 inhibitor S3I-201

dependent variables

Protein expression levels of WASF3, STAT3, pSTAT3 (Tyr 705), and E-cadherin

control variables

Anti-HA, anti-FLAG, and anti-β-Actin antibodies

controls

Positive control: Not specified
Negative control: Not specified

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