1. Volume 56, Issue 1, Pages 83-91 (July 1999)
TGF-βbgr;1 stimulates the release of pre-formed bFGF from renal proximal tubular cells 
Stuart G. Jones, Kimberley Morrisey, John D. Williams, Aled O. Phillips, M.D 
Kidney International 
Volume 56, Issue 1, Pages (July 1999)
DOI: /j x
Copyright © 1999 International Society of Nephrology Terms and Conditions

2. Figure 1
Effect of transforming growth factor-βbgr;1 (TGF-βbgr;1) on basic fibroblast growth factor (bFGF) secretion by primary cultures of human renal proximal tubular cells. Confluent growth-arrested human renal proximal tubular cells were stimulated with recombinant TGF-βbgr;1 (0 to 20 ng/ml) for 24 hours. Supernatant samples were collected, and bFGF was quantitated by ELISA. Data are means ± SD, N = 4; *P < 0.05.
Kidney International  , 83-91DOI: ( /j x)
Copyright © 1999 International Society of Nephrology Terms and Conditions

3. Figure 2
Effect of transforming growth factor-βbgr;1 (TGF-βbgr;1) on basic fibroblast growth factor (bFGF) secretion by HK2 cells. (A) Serum-deprived HK2 cells were stimulated with TGF-βbgr;1 (0 to 10 ng/ml) under serum-free conditions. Supernatant samples were collected after 24 hours, and bFGF was quantitated by ELISA. Data are the means ± SD; for control values N = 25, 1 ng/ml TGF-βbgr;1 N = 11, and 10 ng/ml TGF-βbgr;1 N = 19; *P < (B) Time-dependent effects of TGF-βbgr;1 were determined by the addition of 10 ng/ml of TGF-βbgr;1 to growth-arrested HK2 cells for up to 96 hours. Supernatant bFGF concentration was determined by ELISA. Data are means ± SD, N = 4; *P < 0.05; **P <
Kidney International  , 83-91DOI: ( /j x)
Copyright © 1999 International Society of Nephrology Terms and Conditions

4. Figure 3
Effect of transforming growth factor-βbgr;1 (TGF-βbgr;1) on basic fibroblast growth factor (bFGF) mRNA expression. Serum-deprived HK2 cells were either stimulated with TGF-βbgr;1 (0 to 10 ng/ml) in the absence of serum, and RNA was isolated 24 hours following stimulation (A) or was stimulated with 10 ng/ml TGF-βbgr;1 for up to 96 hours (B). PCR products were separated by electrophoresis on a 3% agarose gel. Scanning densitometry confirmed the lack of induction of bFGF following the addition of TGF-βbgr;1. One representative experiment is shown for both dose- and time-dependent effects of TGF-βbgr;1.
Kidney International  , 83-91DOI: ( /j x)
Copyright © 1999 International Society of Nephrology Terms and Conditions

5. Figure 4
Decreased cell-associated basic fibroblast growth factor (bFGF) concentration following the addition of transforming growth factor-βbgr;1 (TGF-βbgr;1). Serum-deprived cells were stimulated with TGF-βbgr;1 (0 to 10 ng/ml) for 24 hours under serum-free conditions. Cells were detached by treatment with ethylenediaminetetraacetic acid and lyzed as described in the Methods section, and bFGF concentration was determined by enzyme-linked immunosorbent assay. Data are the means ± SD. For control values N = 16, 1 ng/ml TGF-βbgr;1 N = 7, and 10 ng/ml TGF-βbgr;1 N = 13, *P < 0.05.
Kidney International  , 83-91DOI: ( /j x)
Copyright © 1999 International Society of Nephrology Terms and Conditions

6. Figure 5
Effect of transforming growth factor-βbgr;1 (TGF-βbgr;1) on intracellular basic fibroblast growth factor (bFGF). Serum-deprived HK2 cells were stimulated with increasing doses of TGF-βbgr;1 (0 to 10 ng/ml) under serum-free conditions. Alterations in intracellular bFGF were determined by the binding of anti-bFGF antibody to fixed and permeabilized cells. Bound antibody was detected by incubation with alkaline phosphatase-conjugated antimouse antibody (diluted 1/1000 in 0.1% saponin) at room temperature for one hour. Subsequently, para-nitro-phenyl-phosphate (pNPP) was added at a concentration of 1 mg/ml in 10% (wt/vol) diethanolamine buffer containing 0.5 m M MgCl2, pH 9.8. Color was allowed to develop for one hour, and absorbance was read at 410 nm. Results represent the mean OD (410 nm) ± SD. For control values N = 11, 1 ng/ml TGF-βbgr;1 N = 10, and 10 ng/ml TGF-βbgr;1 N = 9, *P < 0.05.
Kidney International  , 83-91DOI: ( /j x)
Copyright © 1999 International Society of Nephrology Terms and Conditions

7. Figure 6
Effect of transforming growth factor-βbgr;1 (TGF-βbgr;1) on matrix associated basic fibroblast growth factor (bFGF). Following stimulation of HK2 cells by TGF-βbgr;1 (0 to 10 ng/ml) under serum-free conditions for 24 hours, cells were detached by treatment with EDTA. The extracellular matrix bound to the plates was subsequently extracted using guanidine hydrochloride. bFGF in the extracted matrix was concentrated using a heparin sepharose column and was subsequently quantitated by ELISA. Data presented are the means ± SD; for control values N = 8, 1 ng/ml TGF-βbgr;1 N = 8, and 10 ng/ml TGF-βbgr;1 N = 9; *P < 0.05.
Kidney International  , 83-91DOI: ( /j x)
Copyright © 1999 International Society of Nephrology Terms and Conditions

8. Figure 7
Methylamine does not affect transforming growth factor-βbgr;1 (TGF-βbgr;1)–induced stimulation of basic fibroblast growth factor (bFGF) secretion. HK2 cells were stimulated with 10 ng/ml TGF-βbgr;1 under serum-free conditions in the presence of increasing doses of methylamine (0 to 20 m M). Supernatant samples were collected after 24 hours, and bFGF concentration was determined by ELISA. Data presented are the means ± SD; for control values N = 20, 10 m M and 20 m M methylamine N = 5.
Kidney International  , 83-91DOI: ( /j x)
Copyright © 1999 International Society of Nephrology Terms and Conditions