fig s1 figure s1: 5t4 expression in b cell development. adapted from microarray data published...
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Fig S1
Figure S1: 5T4 expression in B cell development.Adapted from microarray data published online by van Zelm et al http://franklin.et.tudelft.nl/#bcell [11]
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Fig S2
Figure S2 Expression of 5T4 by B-ALL relapse patients blastsImmunofluorescence staining for 5T4 on DAPI labelled cytospins of bone marrow samples from three relapse patients with pre-B ALL (i-iii) ; magnification x 40, scale bar 5 µm.
SD-1 REH Sup-B15NALM-6 TOM-1
5T4
CXCR4
Sup-B15
TOM-1
A B
Fig S3
C
Figure S3: 5T4 expression by Pre-B ALL cell lines (A) Flow cytometric analyses show the surface expression of 5T4 on a subpopulation of Sup-B15 cells and at low levels on Tom-1 cells, but not by SD-1, REH or Nalm-6 cells. Surface CXCR4 expression is detected on all cells except SD-1. (B) Characteristic punctate 5T4 (green) labelling of cytospins of Sup-B15 and Tom-1 cells (nuclei labelled with DAPI; blue).(C) Immunofluorescence labelling for 5T4 (green), CXCR4 (red ), composite, Z stack and brightfield images for Sup5T4 and Sup cells; bar is 5µM.
5T4 CXCR4 composite Z stack brightfield
5T4 CXCR4 composite Z stack brightfield
Sup5T4
Sup
Fig S4
Sup
Sup5T4
Day 27 ventral images showing assessment of upper and lower abdomen tumour distribution which are presented in Figure 6C as % of total ventral photon count
Fig S5
Bone marrow Ovary
Sup5
T4
5T4
Sup
Figure S5: 5T4 phenotypes are stable after iv challenge and only Sup5T4 is found in the ovaryFlow cytometric analysis representative of bone marrow and ovary cells isolated from mice (n = 4) injected with the Sup-5T4 sublines (5x106 ; i.v.) at day 30 and gated on the mcherry positive leukemic cells. The 5T4 status of the sublines remained unaltered in vivo over time.
Fig S6
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PB
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PB
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NY
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Figure S6: Immunotherapy of Sup5T4 in NSG mice. Ventral images of mice at day 27 after challenge with no treatment, PBMC or PBMC and ANYARA.
Identifier Reference Sup-B15 Sup5T4 Sup
AMEL_1 X X X X
AMEL_2 Y Y Y Y
CSF1PO_1 11 11 11 11
CSF1PO_2 12 12 12 12
D13S317_1 8 8 8 8
D13S317_2 14 14 14 14
D16S539_1 11 11 11 11
D16S539_2 12 12 12 12
D5S818_1 12 12 12 12
D5S818_2 13 13 13 13
D7S820_1 10 10 10 10
D7S820_2 11 10 10 10
TH01_1 6 6 6 6
TH01_2 9.3 9.3 9.3 9.3
TPOX_1 8 8 8 8
TPOX_2 9 9 9 9
vWA_1 15 15 15 15
vWA_2 17 17 17 17
Table S1: Authentication of Sup-B15, Sup & Sup5T4 cells [13]