stat3  (Santa Cruz Biotechnology)

Journal: PLoS ONE

Article Title: Targeting Insulin-Like Growth Factor 1 Receptor Inhibits Pancreatic Cancer Growth and Metastasis

doi: 10.1371/journal.pone.0097016

Figure Lengend Snippet: (A & C): The effect of IGF-1R suppression on ERK and STAT signaling was examined in pancreatic cancer cells. Whole cell lysates were separated by SDS-PAGE and analyzed by Western blot for expression levels of phospho-ERK, ERK, IR-β, phospho-IRS-1, IRS, phospho-STAT3, STAT3, COX-2 and β-actin. (B & D): Representative blots are presented and corresponding densitometric analysis is shown to the right of each image. PS-PANC-1 Scrambled, PI-PANC-1 IGF-1R silenced, HS-HPAC Scrambled, HI-HPAC IGF-1R silenced.

Article Snippet: The following primary antibodies were used in this study: pAKT(sc-101629), AKT(5298), Bcl-2(sc-783), pERK, (sc-101760), ERK (sc-94) and STAT3(H-190)(sc-7179) (Santa Cruz,CA,USA); IGF-1R(3027), Notch 2 (4530P), Snail (3879), E-cadherin (3195), N-cadherin (4061), Zeb (3396), Vimentin (5741), Slug (9585), Bax (2772), Caspase3 (9661), PARP (9542), pPI3K p85(4228), PI3K p85(4292), IR-β (3024), pIRS-1(2388), IRS-1(2382), pSTAT3 (ser727) (4113), COX-2 (4842), pPTEN (9549), pmTOR (2974), mTOR(4517), p-p70s6kinase (9206) and p70s6kinase (9202) (Cell Signaling Technology, (Boston, MA); Caspase8 (ab 25901) (Abcam, Cambridge, MA, USA); β-actin (Sigma Aldrich, (St.Louis, MO, USA).

Techniques: SDS Page, Western Blot, Expressing

Journal: Molecular Therapy. Nucleic Acids

Article Title: LncRNA PVT1 promotes tumorigenesis of glioblastoma by recruiting COPS5 to deubiquitinate and stabilize TRIM24

doi: 10.1016/j.omtn.2021.11.012

Figure Lengend Snippet: PVT1 promotes glioma progression depending on TRIM24 (A) Re-expression of TRIM24 after PVT1 deletion in U251 cells restores cell proliferation. (B) Knockout of TRIM24 after overexpression of PVT1 in LN229 cells impairs cell proliferation. (C and D) Re-expression of TRIM24 after PVT1 deletion in U251 cells restores colony formation ability. (E and F) Knockout of TRIM24 after overexpression of PVT1 in LN229 cells impaired colony formation ability. Scale bars, 10 mm. (G and H) Representative bioluminescence images and H&E-stained images of xenografts' brains with indicated U251 cells stably transfected with PVT1 sh or TRIM24OE. Scale bars, 2 mm. n = 5. (I) WB analysis of the expression of p-STAT3 after transfection with PVT1 sh or TRIM24OE in U251 and LN229 cells. Data are presented as mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: The antibodies used for western blotting were FLAG (no. F3165, Sigma Aldrich), HA (no. 66006-1-Ig, Proteintech), MYC (no. 2276, Cell Signaling Technology), His (no. 2365, Cell Signaling Technology), β-actin (no. 66009-1-Ig, Proteintech), TRIM24 (no. 14208-1-AP, Proteintech), COPS5 (no. SC-13157, Santa Cruz Biotechnology), STAT3 (H-190) (no. SC-7179, Santa Cruz Biotechnology), and phospho-STAT3 (Y705) (D3A7) (no. 9145, Cell Signaling Technology).

Techniques: Expressing, Knock-Out, Over Expression, Staining, Stable Transfection, Transfection

Journal: Molecular Therapy. Nucleic Acids

Article Title: LncRNA PVT1 promotes tumorigenesis of glioblastoma by recruiting COPS5 to deubiquitinate and stabilize TRIM24

doi: 10.1016/j.omtn.2021.11.012

Figure Lengend Snippet: The TRIM24/ PVT1 /COPS5 complex promotes glioma progression through activating STAT3 pathway (A–D) Effects of overexpression of COPS5 and knockdown of PVT1 or knockout of TRIM24 on cell proliferation (A and B) and colony formation ability (C and D). Scale bars, 10 mm (in U251 and U373 cells compared with control). (E and F) Representative bioluminescence images and H&E-stained images of xenografts' brains with indicated U251 cells stably transfected with COPS5OE, PVT1 sh, or TRIM24sg. Scale bars, 2 mm. n = 5. (G) WB analysis of the expression of p-STAT3 after overexpression of COPS5 and knockdown of PVT1 or knockout of TRIM24 in U251 and U373 cells. (H) Schematic model illustrates the mechanism by which PVT1 recruits COPS5 to deubiquitinate and stabilize TRIM24 so as to promote tumorigenesis of GBM. Data are presented as mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: The antibodies used for western blotting were FLAG (no. F3165, Sigma Aldrich), HA (no. 66006-1-Ig, Proteintech), MYC (no. 2276, Cell Signaling Technology), His (no. 2365, Cell Signaling Technology), β-actin (no. 66009-1-Ig, Proteintech), TRIM24 (no. 14208-1-AP, Proteintech), COPS5 (no. SC-13157, Santa Cruz Biotechnology), STAT3 (H-190) (no. SC-7179, Santa Cruz Biotechnology), and phospho-STAT3 (Y705) (D3A7) (no. 9145, Cell Signaling Technology).

Techniques: Over Expression, Knockdown, Knock-Out, Control, Staining, Stable Transfection, Transfection, Expressing