recombinant vegfb  (MedChemExpress)

Journal: International Journal of Molecular Sciences

Article Title: VEGFB Promotes Myoblasts Proliferation and Differentiation through VEGFR1-PI3K/Akt Signaling Pathway

doi: 10.3390/ijms222413352

Figure Lengend Snippet: VEGFB promoted the proliferation of C2C12 cells via VEGFRs signaling. ( A ) Effect of 100 ng/mL of VEGFB and/or 2 µM of axitinib on the proliferation of C2C12 after 48 h culture was determined by CCK8 analysis ( n = 6). ( B ) Effects of 100 ng/mL of VEGFB and/or 2 µM of axitinib on C2C12 proliferation were assessed by using EdU incorporation assay, with the EdU-positive nuclei shown in green. The nuclei were stained with Hoechst, and the scale bar = 200 µm ( n = 3). ( C ) Percentage of EdU-positive cells in panel B. ( D ) Western blot of PCNA and cyclin D1 in C2C12 after 48 h culture. GAPDH was used as loading control. ( E ) Mean ± SEM of immunoblotting bands of PCNA and cyclin D1; the results are expressed as arbitrary units ( n = 6). * p < 0.05 versus control group. n.s = not significant.

Article Snippet: Recombinant mouse VEGFB protein (#293-VE-010) was purchased from R&D systems (Minneapolis, MN, USA).

Techniques: Staining, Western Blot, Control

Journal: International Journal of Molecular Sciences

Article Title: VEGFB Promotes Myoblasts Proliferation and Differentiation through VEGFR1-PI3K/Akt Signaling Pathway

doi: 10.3390/ijms222413352

Figure Lengend Snippet: Knockdown of VEGFR1, but not NRP1, eliminated the promotion of C2C12 proliferation induced by VEGFB. ( A ) Western blot of VEGFR1 and NRP1 in C2C12 after 48 h culture. GAPDH was used as loading control. ( B ) Mean ± SEM of immunoblotting bands of VEGFR1 and NRP1; the results are expressed as arbitrary units ( n = 6). ( C ) Effect of 100 ng/mL of VEGFB and/or siVEGFR1 on the proliferation of C2C12 after 48 h culture was determined by CCK8 analysis ( n = 6). ( D ) Effects of 100 ng/mL of VEGFB and/or siVEGFR1 on C2C12 proliferation were assessed by using EdU incorporation assay ( n = 3). The nuclei were stained with Hoechst. Scale bar = 200 µm. ( E ) Percentage of EdU-positive cells in panel D. ( F ) Western blot analysis of PCNA and cyclin D1 in C2C12 after 48 h culture. β-Tubulin was used as loading control. ( G ) Mean ± SEM of immunoblotting bands of PCNA and cyclin D1; the results are expressed as arbitrary units ( n = 3). ( H ) Effect of 100 ng/mL of VEGFB and/or siNRP1 on the proliferation of C2C12 after 48 h culture was determined by CCK8 analysis ( n = 6). ( I ) Effects of 100 ng/mL of VEGFB and/or siNRP1 on C2C12 proliferation by using EdU incorporation assay ( n = 3). The nuclei were stained with Hoechst. Scale bar = 200 µm. ( J ) Percentage of EdU-positive cells in panel H. * p < 0.05 versus control group. # p < 0.05 versus the siNRP1 group. n.s = not significant. siVEGFR1 and siNRP1, small interfering RNA for VEGFR1 and NRP1, respectively.

Article Snippet: Recombinant mouse VEGFB protein (#293-VE-010) was purchased from R&D systems (Minneapolis, MN, USA).

Techniques: Knockdown, Western Blot, Control, Staining, Small Interfering RNA

Journal: International Journal of Molecular Sciences

Article Title: VEGFB Promotes Myoblasts Proliferation and Differentiation through VEGFR1-PI3K/Akt Signaling Pathway

doi: 10.3390/ijms222413352

Figure Lengend Snippet: VEGFB activated the PI3K/Akt signaling pathway in a VEGFR1-dependent manner, and the inhibition of PI3K/Akt blocked the promotion of C2C12 proliferation induced by VEGFB. ( A ) Western blot analysis of p-PI3K, PI3K, p-Akt, and Akt in C2C12 after 48 h culture. GAPDH was used as loading control. ( B ) Mean ± SEM of immunoblotting bands of p-PI3K, PI3K, p-Akt, and Akt in panel A ( n = 3). ( C ) Western blot of p-PI3K, PI3K, p-Akt, and Akt in C2C12 after 48 h culture. β-Tubulin was used as loading control. ( D ) Mean ± SEM of immunoblotting bands of p-PI3K, PI3K, p-Akt, and Akt in panel C; the results are expressed as arbitrary units ( n = 3). ( E ) Effect of 100 ng/mL of VEGFB and/or 2 µM of Wortmannin on the proliferation of C2C12 after 48 h culture was determined by CCK8 analysis ( n = 6). ( F ) Effects of 100 ng/mL of VEGFB and/or 2 µM of Wortmannin on C2C12 proliferation were assessed by using EdU incorporation assay (n = 3). The nuclei were stained with Hoechst, and the scale bar = 200 µm. ( G ) Percentage of EdU-positive cells in panel F. ( H ) Western blot analysis of PCNA, cyclin D1, p-PI3K, PI3K, p-Akt, and Akt in C2C12 after 48 h culture. GAPDH was used as loading control. ( I ) Mean ± SEM of immunoblotting bands of PCNA, cyclin D1, p-PI3K, PI3K, p-Akt, and Akt; the results are expressed as arbitrary units ( n = 3). * p < 0.05 versus control group. n.s = not significant. siVEGFR1, small interfering RNA for VEGFR1.

Article Snippet: Recombinant mouse VEGFB protein (#293-VE-010) was purchased from R&D systems (Minneapolis, MN, USA).

Techniques: Inhibition, Western Blot, Control, Staining, Small Interfering RNA

Journal: International Journal of Molecular Sciences

Article Title: VEGFB Promotes Myoblasts Proliferation and Differentiation through VEGFR1-PI3K/Akt Signaling Pathway

doi: 10.3390/ijms222413352

Figure Lengend Snippet: VEGFB stimulated the differentiation of C2C12 cells via VEGFRs signaling. ( A ) Effect of 100 ng/mL of VEGFB and/or Axitinib on the differentiation of C2C12 after 5 days of differentiation was determined by immunofluorescence of MyHC ( n = 3). The nuclei were stained with Hoechst, and the scale bar = 200 µm. ( B ) The differentiation index was counted by comparing the MyHC-positive cells to total nuclei. ( C ) Western blot analysis of MyHC, MyoD, and MyoG in C2C12 after 5 days of differentiation. β-Actin was used as loading control. ( D ) Mean ± SEM of immunoblotting bands of MyHC, MyoD, and MyoG; the results are expressed as arbitrary units ( n = 6). * p < 0.05 versus control group. n.s = not significant.

Article Snippet: Recombinant mouse VEGFB protein (#293-VE-010) was purchased from R&D systems (Minneapolis, MN, USA).

Techniques: Immunofluorescence, Staining, Western Blot, Control

Journal: International Journal of Molecular Sciences

Article Title: VEGFB Promotes Myoblasts Proliferation and Differentiation through VEGFR1-PI3K/Akt Signaling Pathway

doi: 10.3390/ijms222413352

Figure Lengend Snippet: Knockdown of VEGFR1, rather than NRP1, was involved in VEGFB-promoted differentiation of C2C12 cells. ( A ) Western blot analysis of VEGFR1 and NRP1 in C2C12 after 5 days of differentiation. β-Actin was used as loading control. ( B ) Mean ± SEM of immunoblotting bands of VEGFR1 and NRP1; the results are expressed as arbitrary units ( n = 6). ( C ) Effect of 100 ng/mL of VEGFB and/or siVEGFR1 on the differentiation of C2C12 after 5 days of differentiation was determined by immunofluorescence of MyHC ( n = 3). The nuclei were stained with Hoechst, and the scale bar = 200 µm. ( D ) The differentiation index was counted by comparing the MyHC-positive cells to total nuclei in panel C. ( E ) Western blot analysis of MyHC, MyoD, and MyoG in C2C12 after 5 days of differentiation. β-Tubulin was used as loading control. ( F ) Mean ± SEM of immunoblotting bands of MyHC, MyoD, and MyoG; the results are expressed as arbitrary units ( n = 3). ( G ) Effect of 100 ng/mL of VEGFB and/or siNRP1 on the differentiation of C2C12 after 5 days of differentiation was determined by immunofluorescence of MyHC ( n = 3). Scale bar = 200 µm ( H ) The differentiation index was counted by comparing the MyHC-positive cells to total nuclei in panel G. The nuclei were stained with Hoechst; * p < 0.05 versus control group. # p < 0.05 versus the siNRP1 group. n.s = not significant. siVEGFR1 and siNRP1, small interfering RNA for VEGFR1 and NRP1, respectively.

Article Snippet: Recombinant mouse VEGFB protein (#293-VE-010) was purchased from R&D systems (Minneapolis, MN, USA).

Techniques: Knockdown, Western Blot, Control, Immunofluorescence, Staining, Small Interfering RNA

Journal: International Journal of Molecular Sciences

Article Title: VEGFB Promotes Myoblasts Proliferation and Differentiation through VEGFR1-PI3K/Akt Signaling Pathway

doi: 10.3390/ijms222413352

Figure Lengend Snippet: VEGFB activated the PI3K/Akt/mTOR signaling pathway in a VEGFR1-dependent manner and the inhibition of PI3K/Akt blocked the promotion of C2C12 differentiation induced by VEGFB. ( A ) Western blot analysis of p-PI3K, PI3K, p-Akt, Akt p-mTOR, mTOR, p-S6, and S6 in C2C12 treated with or without VEGFB and/or axitinib after 5 days of differentiation. ( B ) Mean ± SEM of immunoblotting bands of p-PI3K/PI3K, p-Akt/Akt p-mTOR/mTOR, and p-S6/S6 in panel A. ( C ) Western blot analysis of p-PI3K, PI3K, p-Akt, Akt p-mTOR, mTOR, p-S6, and S6 in C2C12 treated with or without VEGFB and/or siVEGFR1 after 5 days of differentiation. ( D ) Mean ± SEM of immunoblotting bands of p-PI3K/PI3K, p-Akt/Akt, p-mTOR/mTOR, and p-S6/S6 in panel C; the results are expressed as arbitrary units ( n = 3). ( E ) Effect of 100 ng/mL of VEGFB and/or 2 µM of Wortmannin on the differentiation of C2C12 after 5 days of differentiation was determined by immunofluorescence of MyHC ( n = 3). The nuclei were stained with Hoechst and the scale bar = 200 µm. ( F ) The differentiation index was counted by comparing the MyHC-positive cells to total nuclei in panel E. ( G ) Western blot analysis of MyHC, MyoD, MyoG, p-PI3K/PI3K, p-Akt/Akt, p-mTOR/mTOR, and p-S6/S6 in C2C12 after 5 days of differentiation. GAPDH was used as loading control. ( H ) Mean ± SEM of immunoblotting bands of MyHC, MyoD, MyoG, p-PI3K/PI3K, p-Akt/Akt, p-mTOR/mTOR, and p-S6/S6; the results are expressed as arbitrary units ( n = 3). * p < 0.05 versus control group. n.s = not significant. siVEGFR1, small interfering RNA for VEGFR1.

Article Snippet: Recombinant mouse VEGFB protein (#293-VE-010) was purchased from R&D systems (Minneapolis, MN, USA).

Techniques: Inhibition, Western Blot, Immunofluorescence, Staining, Control, Small Interfering RNA

Journal: International Journal of Molecular Sciences

Article Title: VEGFB Promotes Myoblasts Proliferation and Differentiation through VEGFR1-PI3K/Akt Signaling Pathway

doi: 10.3390/ijms222413352

Figure Lengend Snippet: Proposed signaling pathways for VEGFB to promote C2C12 myoblasts proliferation and differentiation.

Article Snippet: Recombinant mouse VEGFB protein (#293-VE-010) was purchased from R&D systems (Minneapolis, MN, USA).

Techniques: Protein-Protein interactions