Supplemental Figure 1
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Fig. S1. Analysis
of Sec31B alternative transcription. Contigs and evidence for alternative
transcription was established by PCR amplification from three separate cDNA
libraries. (A) Sense and anti-sense oligonucleotides used in generating hSec31B
clones in a 5¢ to 3¢ orientation. (B) A graphical representation
of the oligonucleotides used. Sense oligonucleotides are highlighted in cyan
along and numbered above the cDNA for Sec31B; the anti-sense oligonucleotides
are highlighted red and numbered below. (C) Agarose gel analysis of EcoR1 digests of 90 representative amplimers of Sec31B
derived from three independent PCR reactions that were cloned into TA-TOPO
vector. The numbers above each lane represent an ID for independent colonies
picked from a TA-TOPO ligation with various PCR reactions. For all of these
reactions, the same sense and anti-sense primers were used (#52455 and #52458,
respectively). The top band in each lane is the TA-TOPO plasmid. The variation
in the lower bands arises from the considerable alternative splice diversity of
the cDNA for Sec31B.
Supplemental Figure 2
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Fig. S2.
Coincidence of Sec31B-F and VSV-G at ERESs. MDCK-787 cell co-transfected with
ts045-VSV-G-CFP, blocked at 15°C for 3 hours.
Supplemental Figure 3
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Fig. S3.
Coincidence of Sec31B-F, Sec31A, and VSV-G. MDCK-787 cell prepared as in Fig.
S2, stained for Sec31A 15 minutes after moving to permissive temperature.
Supplemental Figure 4
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Fig. S4.
Coincidence of Sec31B with Sec31A. MDCK-787 cell line double stained for Sec31B
(eYFP-Sec31B-F) and Sec31A (also denoted as p137), under various conditions of
fixation, and microscopy (epifluorescence and laser scanning confocal).
Supplemental Figure 5
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Fig. S5.
Relationship of Sec31B to tubulin. MDCK-787 cells transfected with
eCFP-tubulin. Reveals the relationship of microtubules to eYFP-Sec31B-F.
Movie 1
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Movie 1. Dynamics
of Sec31B-F in COS-7 cells. Quicktime® movie of COS-7 cells
transiently transfected with tsO45-VSV-G-CFP (construct 472, cyan). See Fig. 9
for details.
Movie 2
-
Movie 2. Dynamics
of Sec31B-F in COS-7 cells. Quicktime® movie of COS-7 cells
transiently transfected with Sec31B-F-YFP (construct 787, yellow). See Fig. 9
for details.
Movie 3
-
Movie 3. Dynamics
of Sec31B-F in COS-7 cells. Merged Quicktime® movie of COS-7 cells
transiently transfected with tsO45-VSV-G-CFP (construct 472, cyan) and
Sec31B-F-YFP (construct 787, yellow). See Fig. 9 for details.
