Journal: Oncotarget

Article Title: Preclinical assessment of galunisertib (LY2157299 monohydrate), a first-in-class transforming growth factor-β receptor type I inhibitor

doi: 10.18632/oncotarget.23795

Figure Lengend Snippet: Galunisertib inhibits TGFβ-induced pSMAD in 4T1-LP ( A ) or EMT6-LM2 ( B ) cells in vitro . Cells were treated with increasing concentrations of galunisertib (0.20 - 10 μM) then stimulated with TGFβ1 ligand. Following incubation, p- and tSMAD levels were assessed by ELISA. % inhibition of pSMAD2 was calculated as a function of TGFβ1 alone treated cells.

Article Snippet: A standard curve was also added to both ELISA plates (p- and tSMAD plates) using recombinant HIS-pSMAD2 protein (Eli Lilly and Company).

Techniques: In Vitro, Incubation, Enzyme-linked Immunosorbent Assay, Inhibition

Journal: Drug Design, Development and Therapy

Article Title: Treatment with a PPAR-γ Agonist Protects Against Hyperuricemic Nephropathy in a Rat Model

doi: 10.2147/DDDT.S247091

Figure Lengend Snippet: RGTZ abrogates TGF-β-Smad3 signaling in the kidneys of HN rats. ( A ) Kidney tissue lysates were subjected to immunoblot analysis with specific antibodies against TGF-β1 or glyceraldehyde 3-phosphate dehydrogenase (GAPDH). ( B ) Expression levels of TGF-β1 were quantified by densitometry and normalized to GAPDH. ( C ) The kidney tissue lysates were subjected to immunoblot analysis with specific antibodies against p-Smad3, Smad3, or GAPDH. ( D ) Expression levels of p- Smad3 were quantified by densitometry and normalized to Smad3. RGTZ, rosiglitazone. Data are represented as the mean ± SEM. *p < 0.05 vs sham group; #p< 0.05 vs sham + RGTZ group. **p < 0.05 vs HN group.

Article Snippet: Antibodies to Smad3 and p-Smad3 were purchased from Hangzhou HuaAn Biotechnology Co., Ltd. (Hangzhou, China).

Techniques: Western Blot, Expressing

Journal: Aging (Albany NY)

Article Title: Profilin 2 promotes growth, metastasis, and angiogenesis of small cell lung cancer through cancer-derived exosomes

doi: 10.18632/aging.202213

Figure Lengend Snippet: Downstream signaling molecules activated by PFN2 are Smad2/3 in H446 cells and pERK in HUVEC cells. Smad2 and Smad3 expression is increased in H446-OE ( A ) but decreased in H446-KD ( B ). Smad3 inhibitor could impede the function of PFN2 in promoting the proliferation ( P =0.9279, 0.0030, 0.0004, and 0.000 at 6 h, 12 h, 18 h, and 24 h, respectively) ( C ), migration ( P =0.0123) ( D ), and invasion ( P =0.0136) ( E ) of H446 cells. pERK and tERK expression in HUVEC cells co-cultured with H446-OE-exo is upregulated ( F ), whereas the expression in those co-cultured with H446-KD-exo is downregulated ( G ). ERK inhibitor could impede the function of PFN2 in promoting the migration ( P =0.0333) ( H ) and tube formation ability ( P =0.0238) of ECs ( I ).

Article Snippet: The membranes were blocked with 5% bovine serum albumin (BSA) for 1 h before being incubated overnight at 4° C with primary antibodies against PFN2 (Abcam), CD31 (Abcam), TSG101 (Abcam), Alix (Proteintech), Smad2 (Huabio), Smad3 (Immunoway), AKT (CST), p-AKT (CST), ERK (CST), p-ERK (CST), PI3K (CST), or GAPDH (CST).

Techniques: Expressing, Migration, Cell Culture