Journal: Wound Repair and Regeneration

Article Title: Activation of latent transforming growth factor-β1 by nitric oxide in macrophages: Role of soluble guanylate cyclase and MAP kinases

doi: 10.1111/j.1524-475x.2009.00509.x

Figure Lengend Snippet: Figure 2. Nitric oxide induces a dose-dependent increase in ac- tive and latent transforming growth factor-b1 (TGF-b1) protein expression in RAW 264.7 macrophage-like cells. (A) RAW 264.7 cells were treated with 10–1,000 mM S-nitroso-N-acetyl-D,L-pen- icillamine (SNAP) for 24 hours, and the expression of TGF-b1 was analyzed as described in ‘‘Materials and methods.’’ Re- sults shown are the mean SEM from six to seven indepen- dent experiments (np < 0.005 vs. control; nnp < 0.05 vs. 10 mM SNAP). (B) RAW 264.7 cells were treated with 100 mM SNAP or decomposed SNAP (either by heat treatment [T] or pH change) for 24 hours, and the expression of TGF-b1 was analyzed as described in ‘‘Materials and methods.’’ Results shown are the mean SEM. from four independent experiments (np < 0.05 vs. control and decomposed SNAP (pH), nnp < 0.01 vs. decom- posed SNAP (T), #p < 0.05 vs. control and decomposed SNAP (T), ##p < 0.01 vs. decomposed SNAP (pH). (C) RAW 264.7 cells were untreated, treated with 100 mM SNAP, or 100 mM decomposed SNAP. Cell lysates or defined amounts (5, 50, or 100 ng) of recombinant human latency-associated peptide (LAP) were subjected to anti-LAP and b-actin Western blot as described in ‘‘Materials and methods.’’ The experiment is rep- resentative of three; numbers indicate quantification (mean - SEM) of LAP expression normalized for b-actin expression.

Article Snippet: Antibodies to human TGF-b1 and human LAP antibody were purchased from R&D Systems (Minneapolis, MN).

Techniques: Expressing, Control, Recombinant, Western Blot

Journal: Biomedicines

Article Title: Derazantinib Inhibits the Bioactivity of Keloid Fibroblasts via FGFR Signaling

doi: 10.3390/biomedicines11123220

Figure Lengend Snippet: KFs were treated with 2.5 μmol derazantinib or without derazantinib for 48 h. ( A ) The PCR showed that the expression of fibrotic genes was suppressed in the experiment group. ( B – D ) The immunofluorescence staining showed that collagen I, α-SMA, and PAI-1 expression were decreased in the derazantinib group. The length of the scale is 50 μm. ( E , F ) The WB showed that the protein production of a-SMA, collagen I, PAI-1, and FGFR1 in the derazantinib group was suppressed. ( G ) The PCR showed that the expression of PI3K and JNK genes was suppressed in the experiment group. ( H ) The WB showed that the expression of ERK, p-ERK, AKT, TGF-β, p-AKT, and PI3K in the derazantinib group was suppressed. No significant effect was found on the SMAD expression levels. * p < 0.05 compared to control group.

Article Snippet: The PVDF membrane was separately treated with the specific primary antibodies: α-SMA (AF1032, Affinity, Changzhou, China, rabbit), collagen I (66948, CST, USA, mouse), PAI-1 (13801-1-AP, Proteintech, IL, USA, rabbit), FGFR1 (60325-1-Ig, Proteintech, USA, mouse), GAPDH (GB11002, Servicebio, Wuhan, China, rabbit), PI3K (AF5112, Affinity, Changzhou, China, rabbit), p-AKT (AF0016, Affinity, Changzhou, China, rabbit), AKT (BS0115R, Bioss, Wuhan, China, rabbit), p-ERK (AF1015, Affinity, Changzhou, China, rabbit), ERK (AF0155, Affinity, Changzhou, China, rabbit), TGF-β (BA0290, Bosterbio, Beijing, China, rabbit), Smad (6367, Affinity, Changzhou, China, rabbit), and p-Smad (3367, Affinity, Changzhou, China, rabbit).

Techniques: Expressing, Immunofluorescence, Staining, Control