Marker (%)Normal EPCs

Mean ± SM (%)

MMD EPCs Mean

± SM (%)

CD34 26.9 ± 11.5 31.0 ± 6.8

KDR 27.3 ± 5.52 22.4 ± 8.90

CD133 24.5 ± 4.64 12.6 ± 3.93

CD31 99.0 ± 0.9 98.8 ± 0.5

CD45 0.42 ± 1.23 1.53 ± 3.12

CD34 KDR CD133 CD31B

(Scale bar : 50 μm)

CD31 vWF

C D

(Scale bar : 100 μm)

Dil-Ac-LDL uptake

Supplemental Figure I

Normal MMD

(Scale bar : 100 μm)

A 21 days

28 days 35 days

7 days CD45

Normal MMD0

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(Scale bar : 50 μm)

Supplemental Figure II

Se-ries1

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ATRA (-) ATRA 10 μM

Supplemental Figure III(-

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Supplemental Figure I. Phenotypic characterization of endothelial colony forming cells (ECFCs).(A) ECFCs show temporal morphological changes and colonies display a cobblestone morphology. (B) FACS analysis of CD34, KDR, CD133, CD31 and CD45. (C) DiI-labeled Ac-LDL uptake by ECFCs. (D) Immunofluorescence staining of ECFCs labeled with antibodies against CD31 and vWF.  Supplemental Figure II. Retinaldehyde dehydrogenase 2 (RALDH2) knockdown by siRNA transfection in normal endothelial colony forming cells (ECFCs).(A) Visualization of efficient transfection of siRNA into normal ECFCs as green cells. (B) RT-PCR shows effective knockdown of RALDH2 by using two types of siRNA. (C) Similar results from real-time quantitative PCR, demonstrates that the expression of the RALDH2 mRNA is decreased after transfection with siRNA. (D) Gene knockdown was confirmed at the protein level. (N1: normal sample #1, N2: normal sample #2, N3: normal sample #3, N4: normal sample #4) Supplemental Figure III. Effective knockdown retinaldehyde dehydrogenase 2 (RALDH2) and the effect of all-trans retinoic acid (ATRA) treatment on tube formation in human umbilical vein endothelial cell (HUVEC).(A) Normal tube formation capabilities of HUVECs in vitro are inhibited after RALDH2 siRNA was trans-fected. The inhibited tube formation ability of HUVECs after RALDH2 knock down is partially restored after treatment with ATRA. (B) RT-PCR shows effective knock-down of RALDH2 by using 2 types of siRNA in HUVECs. Real-time quantitative PCR also confirms that the expression of RALDH2 mRNA is decreased after transfection with siRNA. (C) The viability and proliferation are not affected by transfection with RALDH2 siRNA.