Journal: International Journal of Molecular Sciences

Article Title: PDGFRα Regulated by miR-34a and FoxO1 Promotes Adipogenesis in Porcine Intramuscular Preadipocytes through Erk Signaling Pathway

doi: 10.3390/ijms18112424

Figure Lengend Snippet: The time-course expression of PDGFRα during porcine IMF preadipocyte differentiation. ( A ) The levels of PDGFRα mRNA by real time qPCR. GAPDH is employed as an internal reference; ( B ) detection of PDGFRα and FABP4 proteins by western blotting. β-tubulin was the loading control; ( C ) the quantify results of protein level. Data represent mean ± SEM, n = 4. * p < 0.05, ** p < 0.01.

Article Snippet: Following primary antibodies were used: PDGFRα (1:300; Boster, Wuhan, China), FABP4 (1:500; Santa Cruz Biotechnology, Dallas, TX, USA), β-actin (1:1000; Santa Cruz Biotechnology, Dallas, TX, USA), β-tubulin (1:500; Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: Expressing, Western Blot, Control

Journal: International Journal of Molecular Sciences

Article Title: PDGFRα Regulated by miR-34a and FoxO1 Promotes Adipogenesis in Porcine Intramuscular Preadipocytes through Erk Signaling Pathway

doi: 10.3390/ijms18112424

Figure Lengend Snippet: Knockdown of modulated- FoxO1 PDGFRα inhibits adipogenesis through Erk signaling pathway. For transfection followed by adipogenic differentiation, cell density must reach 80–85% to ensure that the cells can grow to confluency in two days after transfection. ( A , B ) Real-time qPCR was used to detect PDGFRα , FoxO1 and cell differentiation genes, PPARg and FABP4 after 48 h differentation. GAPDH is employed as an internal reference; ( C ) oil Red O staining. Scale bar = 100 µm; ( D ) western blot analysis of PDGFRα, Erk and p-Erk. β-tubulin was the loading control. ( E – H ) The quantify results of protein level. Data represent mean ± SEM, n = 4. ** p < 0.01.

Article Snippet: Following primary antibodies were used: PDGFRα (1:300; Boster, Wuhan, China), FABP4 (1:500; Santa Cruz Biotechnology, Dallas, TX, USA), β-actin (1:1000; Santa Cruz Biotechnology, Dallas, TX, USA), β-tubulin (1:500; Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: Knockdown, Transfection, Cell Differentiation, Staining, Western Blot, Control

Journal: International Journal of Molecular Sciences

Article Title: PDGFRα Regulated by miR-34a and FoxO1 Promotes Adipogenesis in Porcine Intramuscular Preadipocytes through Erk Signaling Pathway

doi: 10.3390/ijms18112424

Figure Lengend Snippet: Mir-34a targeting PDGFRα represses lipogenesis in IMF preadpocytes. ( A ) The overexpression efficiency of miR-34a after transfecting miR-34a mimics compared with negative control (NC); ( B ) real-time qPCR was used to detect PDGFRα and cell differentiation genes, PPARg and FABP4 after 48 h differentiation. GAPDH is employed as an internal reference; ( C ) oil Red O staining, scale bar = 100 µm; ( D ) western blot analysis of PDGFRα and cell differentiation genes. β-tubulin was the loading control; ( E – I ) the quantify results of protein level. Data are presented as mean ± SEM; n = 4. * p < 0.05, ** p < 0.01.

Article Snippet: Following primary antibodies were used: PDGFRα (1:300; Boster, Wuhan, China), FABP4 (1:500; Santa Cruz Biotechnology, Dallas, TX, USA), β-actin (1:1000; Santa Cruz Biotechnology, Dallas, TX, USA), β-tubulin (1:500; Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: Over Expression, Negative Control, Cell Differentiation, Staining, Western Blot, Control

Journal: International Journal of Molecular Medicine

Article Title: Total flavonoids of Rhizoma Drynariae enhances CD31 hi Emcn hi vessel formation and subsequent bone regeneration in rat models of distraction osteogenesis by activating PDGF-BB/VEGF/RUNX2/OSX signaling axis

doi: 10.3892/ijmm.2022.5167

Figure Lengend Snippet: TFRD promotes CD31 hi Emcn hi vessel formation in angiogenic-osteogenic coupling during distraction osteogenesis via the PDGF-BB/PDGFR-β pathway. (A) Representative immunofluorescence images and (B) quantification of RUNX2, OSX, CD31 and PDGF-BB in the distracted tibias after distraction for 4 weeks. (C) Representative western blotting images and (D) semi-quantitative analyses of PDGF-BB, VEGF, RUNX2, OSX as well as the phosphorylation of AKT and ERK1/2 in the distracted tibias at 4 weeks post-distraction. (E) Quantification of mRNA expression levels of PDGF-BB, VEGF, RUNX2 and OSX. Data represent the mean ± SD. n=3 rats in each group from three independent experiments. *P<0.05 vs. control group; # P<0.05 vs. PDGF-BB-Ab group. TFRD, total flavonoids of Rhizoma drynariae ; PDGF, platelet-derived growth factor; Emcn, endomucin; PDGFR-β, platelet-derived growth factor receptor-β; RUNX2, runt-related transcription factor 2; OSX, Osterix; p-, phosphorylated.

Article Snippet: Subsequently, the membranes were blocked with 5% (w/v) non-fat dry milk in TBS with 0.05% Tween-20 (Beyotime Institute of Biotechnology) at room temperature for 45 min, followed by incubation with primary anti-PDGF-BB (1:500; cat. no. ab21234; Abcam), anti-VEGF (1:500; cat. no. sc-7269; Santa Cruz Biotechnology, Inc.), anti-RUNX2 (1:1,000; cat. no. ab23981; Abcam), anti-OSX (1:1,000; cat. no. ab22552; Abcam), anti-phosphorylated (p)-AKT (1:1,000; cat. no. ab81283; Abcam), p-PDGFR-β (1:2,000; cat. no. AF3132; Affinity Biosciences), PDGFR-β (1:2,000; cat. no. AF6133; Affinity Biosciences), AKT (1:1,000; cat. no. 4691; Cell Signaling Technology, Inc.), ERK (1:2,000; cat. no. AF0155; Affinity Biosciences) and anti-p-ERK1/2 (1:1,000; cat. no. ab214362; Abcam) antibodies at 4°C overnight.

Techniques: Immunofluorescence, Western Blot, Expressing, Derivative Assay

Journal: International Journal of Molecular Medicine

Article Title: Total flavonoids of Rhizoma Drynariae enhances CD31 hi Emcn hi vessel formation and subsequent bone regeneration in rat models of distraction osteogenesis by activating PDGF-BB/VEGF/RUNX2/OSX signaling axis

doi: 10.3892/ijmm.2022.5167

Figure Lengend Snippet: Schematic diagram of the underlying mechanism of TFRD-regulated CD31 hi Emcn hi vessel formation and subsequent bone regeneration during DO. TFRD, total flavonoids of Rhizoma drynariae ; PDGF, platelet-derived growth factor; RUNX2, runt-related transcription factor 2; OSX, Osterix; Emcn, endomucin; PDGFR-β, platelet-derived growth factor receptor-β; DO, distraction osteogenesis.

Article Snippet: Subsequently, the membranes were blocked with 5% (w/v) non-fat dry milk in TBS with 0.05% Tween-20 (Beyotime Institute of Biotechnology) at room temperature for 45 min, followed by incubation with primary anti-PDGF-BB (1:500; cat. no. ab21234; Abcam), anti-VEGF (1:500; cat. no. sc-7269; Santa Cruz Biotechnology, Inc.), anti-RUNX2 (1:1,000; cat. no. ab23981; Abcam), anti-OSX (1:1,000; cat. no. ab22552; Abcam), anti-phosphorylated (p)-AKT (1:1,000; cat. no. ab81283; Abcam), p-PDGFR-β (1:2,000; cat. no. AF3132; Affinity Biosciences), PDGFR-β (1:2,000; cat. no. AF6133; Affinity Biosciences), AKT (1:1,000; cat. no. 4691; Cell Signaling Technology, Inc.), ERK (1:2,000; cat. no. AF0155; Affinity Biosciences) and anti-p-ERK1/2 (1:1,000; cat. no. ab214362; Abcam) antibodies at 4°C overnight.

Techniques: Derivative Assay