goat anti human fgf5  (R&D Systems)

Journal: Stem Cell Research & Therapy

Article Title: Fibroblast growth factor-5 promotes spermatogonial stem cell proliferation via ERK and AKT activation

doi: 10.1186/s13287-019-1139-7

Figure Lengend Snippet: Differentially expressed genes in RNA-Seq were confirmed by Q-PCR. A total of 20 genes, including 10 SCOS Sertoli cells downregulated ( FGF5 , DKK1 , IGF2 , PNP , SEMA3A , CDH1 , SPRY2 , CDK6 , MET and WNK4 ) ( a ) and upregulated ( b ) ones ( SPP1 , TP53I11 , ADRA2A , PLA2G2A , NTN1 , OSCAR , PRRX2 , WFDC1 , NGFR and MT3 ), from 308 DEGs were selected to validate the results of RNA-Seq. Note: OA obstructive azoospermia, SCOS Sertoli cell-only syndrome, SC Sertoli cell. Asterisk indicated statistically significant differences ( P < 0.05)

Article Snippet: Blocking was performed in 1% bovine serum albumin (BSA) for 1 h prior to incubation with primary antibodies, including GATA4 (Santa Cruz, dilution 1:100), WT1 (Santa Cruz, dilution 1:100), and FGF5 (R&D, MAB2371, dilution 1:600).

Techniques: RNA Sequencing

Journal: Stem Cell Research & Therapy

Article Title: Fibroblast growth factor-5 promotes spermatogonial stem cell proliferation via ERK and AKT activation

doi: 10.1186/s13287-019-1139-7

Figure Lengend Snippet: Expression of FGF5 in the human testis of OA and SCOS patients. a RT-PCR showed the expression of FGF5 , FGFR1 , and FGFR2 in freshly isolated human Sertoli cells. ACTB was used as a loading control of total RNA. b Immunocytochemistry showed FGF5 expression in human Sertoli cells of OA and SCOS patients. c Western blot confirmed that expression of FGF5 was decreased at protein level in Sertoli cells from SCOS patients. d IHC revealed the location of FGF5 expression in the human testis (scale bar = 10 μm). Asterisk indicated the location of FGF5 expression in the human testicular section

Article Snippet: Blocking was performed in 1% bovine serum albumin (BSA) for 1 h prior to incubation with primary antibodies, including GATA4 (Santa Cruz, dilution 1:100), WT1 (Santa Cruz, dilution 1:100), and FGF5 (R&D, MAB2371, dilution 1:600).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Isolation, Control, Immunocytochemistry, Western Blot

Journal: Stem Cell Research & Therapy

Article Title: Fibroblast growth factor-5 promotes spermatogonial stem cell proliferation via ERK and AKT activation

doi: 10.1186/s13287-019-1139-7

Figure Lengend Snippet: The effect of FGF5 in the proliferation of C18-4 (mouse SSCs line). a In CCK-8 assay, the growth of C18-4 cell line with various concentrations of FGF5 for 3 days (0, 1, 5, 25, 100, 200, and 400 ng/ml). b CCK-8 showed the growth activity of C18-4 cell line after treatment of FGF5 (100 ng/ml) for 1–5 days. c EdU incorporation assay revealed the proliferation activity after treatment of FGF5 (100 ng/ml) for 3 days. Asterisk indicated statistically significant differences ( P < 0.05) between treatment of FGF5 and with the control

Article Snippet: Blocking was performed in 1% bovine serum albumin (BSA) for 1 h prior to incubation with primary antibodies, including GATA4 (Santa Cruz, dilution 1:100), WT1 (Santa Cruz, dilution 1:100), and FGF5 (R&D, MAB2371, dilution 1:600).

Techniques: CCK-8 Assay, Activity Assay, Control

Journal: Stem Cell Research & Therapy

Article Title: Fibroblast growth factor-5 promotes spermatogonial stem cell proliferation via ERK and AKT activation

doi: 10.1186/s13287-019-1139-7

Figure Lengend Snippet: Phosphoprotein profiles in C18-4 cells after treatment of 100 ng/ml FGF5 in 5 and 15 min. a Distinct phosphoprotein profiles in C18-4 cells after 100 ng/ml FGF5 in 5- and 15-min treatment compared with the control. b KEGG informatics analysis showed the pathway enrichment of differentially phosphorylated proteins after treatment of 100 ng/ml FGF5 in 5 and 15 min compared with the control

Article Snippet: Blocking was performed in 1% bovine serum albumin (BSA) for 1 h prior to incubation with primary antibodies, including GATA4 (Santa Cruz, dilution 1:100), WT1 (Santa Cruz, dilution 1:100), and FGF5 (R&D, MAB2371, dilution 1:600).

Techniques: Control

Journal: Stem Cell Research & Therapy

Article Title: Fibroblast growth factor-5 promotes spermatogonial stem cell proliferation via ERK and AKT activation

doi: 10.1186/s13287-019-1139-7

Figure Lengend Snippet: The expression changes of ERK, AKT, p-CREB, c-fos, STAT3, PCNA, Cyclin A2, and Cyclin E after treatment of FGF5. a , b Western blot revealed the expression changes of p-ERK and p-AKT after 100-ng/ml FGF5 treatment. ERK, AKT, and ACTB were used as loading controls. c The expression changes of p-CREB, c-fos, and STAT3 after treatment of FGF5 for 5 and 15 min. d The expression changes of PCNA, Cyclin A2, and Cyclin E after 100 ng/ml FGF5 treatment for 6 and 12 h

Article Snippet: Blocking was performed in 1% bovine serum albumin (BSA) for 1 h prior to incubation with primary antibodies, including GATA4 (Santa Cruz, dilution 1:100), WT1 (Santa Cruz, dilution 1:100), and FGF5 (R&D, MAB2371, dilution 1:600).

Techniques: Expressing, Western Blot

Journal: Stem Cell Research & Therapy

Article Title: Fibroblast growth factor-5 promotes spermatogonial stem cell proliferation via ERK and AKT activation

doi: 10.1186/s13287-019-1139-7

Figure Lengend Snippet: Differentially expressed genes in RNA-Seq were confirmed by Q-PCR. A total of 20 genes, including 10 SCOS Sertoli cells downregulated ( FGF5 , DKK1 , IGF2 , PNP , SEMA3A , CDH1 , SPRY2 , CDK6 , MET and WNK4 ) ( a ) and upregulated ( b ) ones ( SPP1 , TP53I11 , ADRA2A , PLA2G2A , NTN1 , OSCAR , PRRX2 , WFDC1 , NGFR and MT3 ), from 308 DEGs were selected to validate the results of RNA-Seq. Note: OA obstructive azoospermia, SCOS Sertoli cell-only syndrome, SC Sertoli cell. Asterisk indicated statistically significant differences ( P < 0.05)

Article Snippet: After blocking with 5% milk, membranes were incubated with antibodies against FGF5 (R&D, MAB2371, dilution 1:2000), p-ERK1/2 (CST, 4370, dilution 1:2000), ERK1/2 (CST, 4695, dilution 1:1000), p-AKT (CST, 4060, dilution 1:2000), AKT (CST, 4691, dilution 1:1000), p-CREB (Santa Cruz, sc-81486, dilution 1:1000), c-fos (Santa Cruz, sc-447, dilution 1:600), STAT3 (Santa Cruz, sc-8019, dilution 1:200), PCNA (Santa Cruz, sc-25280, dilution 1:600), Cyclin A2 (Proteintech, 18202-1-AP, dilution 1:2000), Cyclin E1 (Proteintech, 11554-1-AP, dilution 1:2000), or ACTB (beta-actin) (Santa Cruz, sc-47778, dilution 1:500) at 4 °C overnight.

Techniques: RNA Sequencing

Journal: Stem Cell Research & Therapy

Article Title: Fibroblast growth factor-5 promotes spermatogonial stem cell proliferation via ERK and AKT activation

doi: 10.1186/s13287-019-1139-7

Figure Lengend Snippet: Expression of FGF5 in the human testis of OA and SCOS patients. a RT-PCR showed the expression of FGF5 , FGFR1 , and FGFR2 in freshly isolated human Sertoli cells. ACTB was used as a loading control of total RNA. b Immunocytochemistry showed FGF5 expression in human Sertoli cells of OA and SCOS patients. c Western blot confirmed that expression of FGF5 was decreased at protein level in Sertoli cells from SCOS patients. d IHC revealed the location of FGF5 expression in the human testis (scale bar = 10 μm). Asterisk indicated the location of FGF5 expression in the human testicular section

Article Snippet: After blocking with 5% milk, membranes were incubated with antibodies against FGF5 (R&D, MAB2371, dilution 1:2000), p-ERK1/2 (CST, 4370, dilution 1:2000), ERK1/2 (CST, 4695, dilution 1:1000), p-AKT (CST, 4060, dilution 1:2000), AKT (CST, 4691, dilution 1:1000), p-CREB (Santa Cruz, sc-81486, dilution 1:1000), c-fos (Santa Cruz, sc-447, dilution 1:600), STAT3 (Santa Cruz, sc-8019, dilution 1:200), PCNA (Santa Cruz, sc-25280, dilution 1:600), Cyclin A2 (Proteintech, 18202-1-AP, dilution 1:2000), Cyclin E1 (Proteintech, 11554-1-AP, dilution 1:2000), or ACTB (beta-actin) (Santa Cruz, sc-47778, dilution 1:500) at 4 °C overnight.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Isolation, Control, Immunocytochemistry, Western Blot

Journal: Stem Cell Research & Therapy

Article Title: Fibroblast growth factor-5 promotes spermatogonial stem cell proliferation via ERK and AKT activation

doi: 10.1186/s13287-019-1139-7

Figure Lengend Snippet: The effect of FGF5 in the proliferation of C18-4 (mouse SSCs line). a In CCK-8 assay, the growth of C18-4 cell line with various concentrations of FGF5 for 3 days (0, 1, 5, 25, 100, 200, and 400 ng/ml). b CCK-8 showed the growth activity of C18-4 cell line after treatment of FGF5 (100 ng/ml) for 1–5 days. c EdU incorporation assay revealed the proliferation activity after treatment of FGF5 (100 ng/ml) for 3 days. Asterisk indicated statistically significant differences ( P < 0.05) between treatment of FGF5 and with the control

Article Snippet: After blocking with 5% milk, membranes were incubated with antibodies against FGF5 (R&D, MAB2371, dilution 1:2000), p-ERK1/2 (CST, 4370, dilution 1:2000), ERK1/2 (CST, 4695, dilution 1:1000), p-AKT (CST, 4060, dilution 1:2000), AKT (CST, 4691, dilution 1:1000), p-CREB (Santa Cruz, sc-81486, dilution 1:1000), c-fos (Santa Cruz, sc-447, dilution 1:600), STAT3 (Santa Cruz, sc-8019, dilution 1:200), PCNA (Santa Cruz, sc-25280, dilution 1:600), Cyclin A2 (Proteintech, 18202-1-AP, dilution 1:2000), Cyclin E1 (Proteintech, 11554-1-AP, dilution 1:2000), or ACTB (beta-actin) (Santa Cruz, sc-47778, dilution 1:500) at 4 °C overnight.

Techniques: CCK-8 Assay, Activity Assay, Control

Journal: Stem Cell Research & Therapy

Article Title: Fibroblast growth factor-5 promotes spermatogonial stem cell proliferation via ERK and AKT activation

doi: 10.1186/s13287-019-1139-7

Figure Lengend Snippet: Phosphoprotein profiles in C18-4 cells after treatment of 100 ng/ml FGF5 in 5 and 15 min. a Distinct phosphoprotein profiles in C18-4 cells after 100 ng/ml FGF5 in 5- and 15-min treatment compared with the control. b KEGG informatics analysis showed the pathway enrichment of differentially phosphorylated proteins after treatment of 100 ng/ml FGF5 in 5 and 15 min compared with the control

Article Snippet: After blocking with 5% milk, membranes were incubated with antibodies against FGF5 (R&D, MAB2371, dilution 1:2000), p-ERK1/2 (CST, 4370, dilution 1:2000), ERK1/2 (CST, 4695, dilution 1:1000), p-AKT (CST, 4060, dilution 1:2000), AKT (CST, 4691, dilution 1:1000), p-CREB (Santa Cruz, sc-81486, dilution 1:1000), c-fos (Santa Cruz, sc-447, dilution 1:600), STAT3 (Santa Cruz, sc-8019, dilution 1:200), PCNA (Santa Cruz, sc-25280, dilution 1:600), Cyclin A2 (Proteintech, 18202-1-AP, dilution 1:2000), Cyclin E1 (Proteintech, 11554-1-AP, dilution 1:2000), or ACTB (beta-actin) (Santa Cruz, sc-47778, dilution 1:500) at 4 °C overnight.

Techniques: Control

Journal: Stem Cell Research & Therapy

Article Title: Fibroblast growth factor-5 promotes spermatogonial stem cell proliferation via ERK and AKT activation

doi: 10.1186/s13287-019-1139-7

Figure Lengend Snippet: The expression changes of ERK, AKT, p-CREB, c-fos, STAT3, PCNA, Cyclin A2, and Cyclin E after treatment of FGF5. a , b Western blot revealed the expression changes of p-ERK and p-AKT after 100-ng/ml FGF5 treatment. ERK, AKT, and ACTB were used as loading controls. c The expression changes of p-CREB, c-fos, and STAT3 after treatment of FGF5 for 5 and 15 min. d The expression changes of PCNA, Cyclin A2, and Cyclin E after 100 ng/ml FGF5 treatment for 6 and 12 h

Article Snippet: After blocking with 5% milk, membranes were incubated with antibodies against FGF5 (R&D, MAB2371, dilution 1:2000), p-ERK1/2 (CST, 4370, dilution 1:2000), ERK1/2 (CST, 4695, dilution 1:1000), p-AKT (CST, 4060, dilution 1:2000), AKT (CST, 4691, dilution 1:1000), p-CREB (Santa Cruz, sc-81486, dilution 1:1000), c-fos (Santa Cruz, sc-447, dilution 1:600), STAT3 (Santa Cruz, sc-8019, dilution 1:200), PCNA (Santa Cruz, sc-25280, dilution 1:600), Cyclin A2 (Proteintech, 18202-1-AP, dilution 1:2000), Cyclin E1 (Proteintech, 11554-1-AP, dilution 1:2000), or ACTB (beta-actin) (Santa Cruz, sc-47778, dilution 1:500) at 4 °C overnight.

Techniques: Expressing, Western Blot