doxycycline dox induction  (LKT Laboratories)

Journal: The Journal of Clinical Investigation

Article Title: Androgen deprivation–mediated activation of AKT is enhanced in prostate cancer with TMPRSS2:ERG fusion

doi: 10.1172/JCI192368

Figure Lengend Snippet: ( A ) H19 and IGF2 expression in mice with long-term DOX induction versus after DOX withdrawal. ( B ) H19, IGF2, and ERG expression in mouse prostate expressing probasin-ERG transgenes versus WT (from ref. ). ( C ) Correlation between ERG and H19 mRNA in TCGA primary PCa. ( D ) Correlation between ERG and H19 mRNA in T:E fusion–positive tumors in TCGA primary PCa. ( E ) Correlation between ERG and H19 mRNA in T:E fusion–negative tumors in TCGA primary PCa. Correlations were assessed in cBioPortal.

Article Snippet: RNA from mouse prostate prior to and after DOX induction was analyzed by hybridization on Affymetrix Mouse Gene 1.0 ST arrays.

Techniques: Expressing

Journal: bioRxiv

Article Title: Androgen deprivation-mediated activation of AKT is enhanced in prostate cancer with TMPRSS2:ERG fusion

doi: 10.1101/2024.08.26.609679

Figure Lengend Snippet: (A) H19 and IGF2 expression in mice with long-term DOX induction versus after DOX withdrawal. (B) H19, IGF2, and ERG expression in mouse prostate expressing probasin-ERG transgenes versus wild-type (from reference 13). (C) Correlation between ERG and H19 mRNA in TCGA primary PCa. (D) Correlation between ERG and H19 mRNA in T:E fusion positive tumors in TCGA primary PCa. (E) Correlation between ERG and H19 mRNA in T:E fusion negative tumors in TCGA primary PCa. Correlations were assessed in cBioPortal.

Article Snippet: RNA from mouse prostate prior to and post DOX induction was analyzed by hybridization on Affymetrix Mouse Gene 1.0 ST arrays.

Techniques: Expressing

Journal: Scientific reports

Article Title: Transcription factor FoxO1 regulates myoepithelial cell diversity and growth.

doi: 10.1038/s41598-024-51619-1

Figure Lengend Snippet: Figure 2. Overexpression of FoxO1 in ME cells. (A) Scatter plot of CD49f (x-axis) and EpCAM (y-axis). The cells were isolated from SMG in TP53 mutant female mice (n = 4) and analyzed by flow cytometry. EpCAMlowCD49fhigh-cells were sorted as ME cells (6.5%). (B) A schematic for integration of PiggyBac transposon vector plasmid. The Tet-On inducible gene expression system was used. FoxO1 expression was induced by doxycycline (Dox). (C) mCherry fluorescence merged with phase contrast in MEPB-FoxO1 cells treated with and without Dox (2 µg/mL) for 48 h. (D) Expression of FoxO1 mRNA in MEPB-FoxO1 cells treated with and without Dox for 24 h. *P < 0.05. n = 3. (E) Immunoblotting for FoxO1, αSMA, Krt14, Krt5, and β-actin in MEPB-FoxO1 cells treated with and without Dox for 72 h. (F) FoxO1 luciferase assay in the presence of FoxO1 inhibitor (Inh.; AS1842856) at the indicated concentrations. pGL4 luciferase reporter vector (upper) was constructed to include three FoxO1-binding elements (daf16:TTGTTTA and mdaf16:TTGCTTA). FoxO1 transcriptional activity was measured. pRL-TK was used as internal control. The Renilla luciferase normalized the firefly luciferase. #P < 0.05 vs. control (Ctrl). *P < 0.05 vs. Dox. n = 5. (G) Expression of αSMA mRNA in ME cells treated with and without FoxO1 inhibitor (Inh.; AS1842856, 1 μM) for 72 h. *P < 0.05. n = 3. (H) Expression of FoxO1 and αSMA mRNA in siRNA-mediated knockdown of FoxO1 (siFoxO1) or control (si Ctrl) in ME cells. *P < 0.05. n = 3. (I) Immunoblotting for NF-κB/p65 and phospho-NF-κB/p65 in MEPB-FoxO1 cells treated with and without Dox at the indicated time-points. The signal intensity of phospho-NF-κB/p65 was normalized to that of NF-κB/p65 (ratio). All data were representative of three independent experiments. See also Supplementary Figs. S2 and S3.

Article Snippet: The PiggyBac plasmid carried a tetracycline response element to drive doxycycline (Dox) induction (2 μg/mL; LKT Labs, St. Paul, MN).

Techniques: Over Expression, Isolation, Mutagenesis, Flow Cytometry, Plasmid Preparation, Gene Expression, Expressing, Fluorescence, Western Blot, Luciferase, Construct, Binding Assay, Activity Assay, Control, Knockdown