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1
Supplemental Figure 1. Specific siRNA treatments
suppress the mRNA expression of each PLC isoform
in MIN-6 cells Non-targeting (NT) siRNA (open
bars n5) or siRNA specific for each PLC
isoform (closed bar n5) was applied to MIN-6
cells. After a 48 hr incubation, cells were
subjected to RT-PCR analysis. PLC expression
levels in MIN-6 cells were quantified and
normalized relative to ß-actin mRNA levels. P
lt0.05, P lt0.01 versus NT siRNA-transfected
MIN-6 cells assessed by unpaired t test. Data
are presented as means SEM.
PLCß2
PLCß1
PLCß3



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PLCß1
NT
siRNA
NT
siRNA
NT
siRNA
PLCß2
PLCß3
PLC?2
PLCd1
PLC?1



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PLC?1
NT
siRNA
NT
siRNA
NT
siRNA
PLC?2
PLCd1
2
Supplemental Figure 2. Specific siRNA treatments
suppress the protein levels of PLC-ß1 and PLC-?1
isoforms in MIN-6 cells Non-targeting (NT) siRNA
or siRNA specific for each PLC-ß1 and PLC-?1
isoforms were applied to MIN-6 cells. After a 48
hr incubation, immunoblot analysis was performed
with MIN-6 cells lysate, using a monoclonal anti-
PLCß1 antibody (sc-5291, Santa Cruz) or a
polyclonal anti- PLC?1 antibody (2822, Cell
Signaling).
3
Supplemental Table 1. The oligonucleotide primers
used for RT-PCR.
4
Supplemental Table 2. Effects of several
pretreatments on insulin content of MIN-6 cells
and isolated islets Insulin contents of MIN-6
cells (1-5106 cells/well) and isolated islets
(10 islets/well) used in the experiments are
indicated. Left column shows the Figure numbers
corresponding to each pretreatments. Data are
presented as means SEM.
Insulin content (ng)
Pretreatments
Fig.
IL-6 (-) IL-6 ()
IL-6 1200 pg/ml (isolated islets) 426.5 25.4 450.7 17.4
IL-6 600 pg/ml (isolated islets) 426.5 25.4 479.5 34.5
vehicle (isolated islets) 241.7 43.1 208.6 24.0
U-73122 (isolated islets) 198.5 57.3 171.7 35.0
IL-6 1200 pg/ml 6 hr (MIN-6) 3025.0 34.1 3094.2 181.1
IL-6 1200 pg/ml 24 hr (MIN-6) 3277.3 63.4 3190.1 165.6
IL-6 1200 pg/ml 48 hr (MIN-6) 2909.9 73.0 2776.7 62.3
NT siRNA (MIN-6) 4683.9 100.6 5238.1 62.0
IL-6R siRNA (MIN-6) 4032.1 198.9 4844.1 196.1
Vehicle (MIN-6) 1342.5 175.4 1232.3 195.9
U-73122 (MIN-6) 1529.1 39.8 1426.3 21.2
Neomycin (MIN-6) 1645.9 41.4 1585.9 24.3
NT siRNA (MIN-6) 2702.0 97.7 2816.2 129.0
PLC ß1 siRNA (MIN-6) 2899.0 18.9 3171.5 19.0
PLC ß2 siRNA (MIN-6) 2556.5 72.3 2915.4 169.3
PLC ß3 siRNA (MIN-6) 2389.7 36.8 2594.7 78.2
PLC ?1 siRNA (MIN-6) 3004.8 71.6 2875.3 56.7
PLC ?2 siRNA (MIN-6) 2378.7 92.0 2664.1 100.9
PLC d1 siRNA (MIN-6) 2881.5 65.8 3138.3 43.5
Vehicle (MIN-6) 3700.5 75.5 3721.5 51.6
H-89 (MIN-6) 3442.3 74.3 3700.0 96.4
Vehicle (MIN-6) 1506.2 24.1 1461.4 35.9
Xestospongin C (MIN-6) 762.3 14.9 675.7 139.7