Supplementary Figure 1: Technical reproducibility of CTC enumeration and
PTEN status. (a) CTC enumeration in duplicate slides from 89 CRPC samples. Each
dot represents a patient sample with CTC counts from slide 1 on the X-axis, and
CTC counts from slide 2 on the Y-axis. CTC=0 were converted to 0.5 to enable log2
graph (b) Comparison of PTEN status evaluated in 2 replicate slides. N=10 patients
with ≥ 4 CTCs per slide, sufficient for independent PTEN evaluation in 1 slide.
Supplementary Figure 2:
CEP and PTEN genotypes in WBC from healthy
donors. (a) Mean frequencies of CEP10 and PTEN genotypes in WBC. x, number of
CEP10 signals per cell, y, number of PTEN signals per cell. N= 40 cells per donor,
10 donors.
Supplementary Figure 3: Representative images of concordant PTEN status in
matched CTCs, archived and fresh tumor tissue specimens. Upper panel:
concordant HO loss in CTCs, archived and fresh tissue; middle panel: concordant
HO/HE loss mix in CTCs and fresh tissue; lower panel: concordant PTEN Nondeleted in CTCs, archived and fresh tissue. Dotted circles demarcate CTCs. Blue
stain=DAPI, red signals=PTEN, green signals=CEP10.
Supplementary Figure 4: Comparison of PTEN status in CTCs (FISH) with PTEN
status in tissue specimens (IHC). (a) PTEN status in CTCs vs. fresh tissue. (b).
PTEN status in CTCs vs. archived tissue. Bottom, PTEN concordance in CTCs vs.
fresh and archived biopsies. ** two patients with heterogeneous PTEN status in
archival were not included in the analysis.
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Supplementary Figure 5: Comparison of CTC enumeration in CellSearch vs.
EPIC. (a) CellSearch and EPIC CTC enumeration are indicated as CTCs counts/7.5
ml. Each bar represents EPIC (red) and CellSearch (black) in replicate samples
(n=69) from 66 patients (b) IF (top) and FISH (bottom) images of CTCs from patients
5039 (2-cell clusters in i and ii) and 5059 (single CTCS IN iii and iv), which showed
high EPIC CTCs counts but low Veridex CTC count.
Supplementary Figure 6: Representative images of AR staining in CTCs
Upper panel: nuclear localization of AR in patient 5052 CTC; Middle panel:
cytoplasmic localization of AR in patient 5052 CTC; Lower panel: mixed localization
of AR in patient 5052 CTC. Blue = DAPI, red = CK, green = CD45, white = AR.
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