Alex Limanni, M. D. , Timothy Fleming, Ph. D. , Rodolfo Molina, M. D

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Presentation transcript:

Expression of genes for platelet-derived growth factor in adult human venous endothelium Alex Limanni, M.D. *, Timothy Fleming, Ph.D., Rodolfo Molina, M.D., Howard Hufnagel, B.S., Robert E. Cunningham, M.S., David F. Cruess, Ph.D., John B. Sharefkin, M.D. Journal of Vascular Surgery Volume 7, Issue 1, Pages 10-20 (January 1988) DOI: 10.1016/0741-5214(88)90374-6 Copyright © 1988 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 1 A. Northern blot for RNA hybridizing to a probe with a sequence encoding the B chain of PDGF. Five micrograms of cytoplasmic RNA were loaded per lane. Lane A: bovine aortic ECs. Lane B: human dermal fibroblasts. All other lanes represent four AHSVEC lines (C,D,E, and F) subjected to either 48 hours of withdrawal of retinal extract and heparin (lanes C1, D1, E1, and F1) or 48 hours of continued use of retinal extract and heparin up to the time of RNA extraction (lanes C2, D2, E2, and F2). The accompanying bar graph represents scanning densitometric signal strengths for each lane divided by the number of cells counted in the two T-150 flasks used for each lane at the end of the 48-hour treatment period (upper bar graph) or a similar plot of signal strength divided by the total amount of cytoplasmic RNA in micrograms obtained from the same two flasks (lower bar graph). B, Northern blot for RNA hybridizing to a probe for the human PDGF A chain cloned from a human glioma cell line. Five micrograms of cytoplasmic RNA were loaded per lane. Lane designations for cell lines and treatment groups are the same as for A. Bar graphs of scanning densitometer signal per cell and per microgram total RNA are the same as in A, except that one AHSVEC line signal (lane A) could not be scanned because of nonspecific background film darkening near the A chain bands. Journal of Vascular Surgery 1988 7, 10-20DOI: (10.1016/0741-5214(88)90374-6) Copyright © 1988 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 2 Effect on tritiated thymidine uptake by postconfluent NIH 3T3 cells of no treatment (A), 3 ng/ml purified human platelet-derived PDGF (B), purified PDGF plus monospecific anti-human PDGF antibody (C), addition of 50% vol/vol of serum-free medium exposed for 48 hours to confluent sixth passage AHSVECs, and conditioned medium effect plus the same antibody used for (B) (E). Bars show mean and standard deviation of beta counts for triplicate 16 mm wells. Journal of Vascular Surgery 1988 7, 10-20DOI: (10.1016/0741-5214(88)90374-6) Copyright © 1988 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 3 Flow cytometric histograms of relative cell number vs. DNA content for cultures of HDF in medium 199 with 15% fetal bovine serum (A), with bovine aortic ECs in Dulbecco's modified Eagle's medium with 10% calf serum (B), an AHSVEC culture 48 hours after heparin and retinal extract withdrawal (C), and replicate flasks of the same AHSVEC culture after 48-hour maintenance of retinal extract and heparin treatment (D). Journal of Vascular Surgery 1988 7, 10-20DOI: (10.1016/0741-5214(88)90374-6) Copyright © 1988 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions