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Legends for supplementary figures Fig s1. A: Western blot showing the suppression of SRp20 expression by Dox induction of SRp20 siRNAs in SKOV3 sublines. B: Cell growth curve. Shown are the results of three independent experiments. Error bars represent standard error. C: Left panel, sample micrographs of Hoechst 33342-stained nuclei of SKOV3 subline cells. Arrow indicates the typical apoptotic cells. Right panel, Quantitation of apoptotic cells. Fig s2. A: Western blot showing the suppression of SRp20 expression by Dox induction of SRp20 siRNAs in IGROV1 sublines. B: Cell growth curve of IGROV1 sublines. C: Left panel, sample micrographs of Hoechst 33342-stained nuclei of IGROV1 subline cells. Arrow indicates the typical apoptotic cells. Right panel, Quantitation of apoptotic cells.
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Legends for supplementary figures Fig s1. A: Western blot showing the suppression of SRp20 expression by Dox induction of SRp20 siRNAs in SKOV3 sublines.

Dec 18, 2015

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Coleen Glenn
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Page 1: Legends for supplementary figures Fig s1. A: Western blot showing the suppression of SRp20 expression by Dox induction of SRp20 siRNAs in SKOV3 sublines.

Legends for supplementary figures

Fig s1. A: Western blot showing the suppression of SRp20 expression

by Dox induction of SRp20 siRNAs in SKOV3 sublines. B: Cell growth

curve. Shown are the results of three independent experiments. Error

bars represent standard error. C: Left panel, sample micrographs of

Hoechst 33342-stained nuclei of SKOV3 subline cells. Arrow indicates

the typical apoptotic cells. Right panel, Quantitation of apoptotic cells.

Fig s2. A: Western blot showing the suppression of SRp20 expression

by Dox induction of SRp20 siRNAs in IGROV1 sublines. B: Cell growth

curve of IGROV1 sublines. C: Left panel, sample micrographs of

Hoechst 33342-stained nuclei of IGROV1 subline cells. Arrow indicates

the typical apoptotic cells. Right panel, Quantitation of apoptotic cells.

Page 2: Legends for supplementary figures Fig s1. A: Western blot showing the suppression of SRp20 expression by Dox induction of SRp20 siRNAs in SKOV3 sublines.

Dox

SKOV3β-actin

SRp20

SRp20si1 SRp20si2- + - +

LUCsi- +

Fig s1

A

B

C

SKOV3/LUCsi

SKOV3/SRp20si1

SKOV3/SRp20si2

Without Dox WithDox

% Knockdown 67 91

Page 3: Legends for supplementary figures Fig s1. A: Western blot showing the suppression of SRp20 expression by Dox induction of SRp20 siRNAs in SKOV3 sublines.

SRp20si1 SRp20si2LUCsi

- + - +Dox - +

β-actin

SRp20IGROV1

Fig s2

A

B

CWithout Dox With Dox

IGROV1/LUCsi

IGROV1/SRp20si1

IGROV1/SRp20si2

% Knockdown 47 79

Page 4: Legends for supplementary figures Fig s1. A: Western blot showing the suppression of SRp20 expression by Dox induction of SRp20 siRNAs in SKOV3 sublines.

† Evaluable cases require at least two satisfactory cores with staining for either PTB or SRp20.

Disease Status

Satisfactory Cores per Case Evaluable

Cases †TotalCases

Histologic Subtype for Evaluable Cases

0 1 2 3 Mucinous Serous Clear Cell EndometrioidPoorly

Differentiated

PTB staining

Benign 12 13 14 3 17 42 15 2 0 0 0

Borderline/LMP 0 12 18 31 49 61 25 23 0 1 0

Invasive 1 2 11 56 67 70 26 9 30 0 2

Total 13 27 43 90 133 173 66 34 30 1 2

SRp20 staining

Benign 23 11 5 3 8 42 7 1 0 0 0

Borderline/LMP 2 14 15 30 45 61 22 22 0 1 0

Invasive 2 4 6 58 64 70 25 9 28 0 2

Total 27 29 26 91 117 173 54 32 28 1 2

Table s1. Distribution of Evaluable and Total Cases by Disease Status, Number of Satisfactory Cores and Histologic Subtype in the Disease Status TMA Stained for PTB or SRp20.

Page 5: Legends for supplementary figures Fig s1. A: Western blot showing the suppression of SRp20 expression by Dox induction of SRp20 siRNAs in SKOV3 sublines.

Disease status

PTB † SRp20 ‡

All Negative

MixedAll

PositiveTotal

All Negative

MixedAll

PositiveTotal

Benign 7 (46.7) 5 (33.3) 3 (20.0) 15 1 (14.3) 3 (42.9) 3 (42.9) 7

Borderline/LMP 8 (32.0) 7 (28.0) 10 (40.0) 25 0 (0) 9 (40.9) 13 (59.1) 22

Invasive 2 (7.7) 5 (19.2) 19 (73.1) 26 0 (0) 2 (8.0) 23 (92.0) 25

Total 17 17 32 66 1 14 39 54

Table s2. Differential expression of PTB or SRp20 in the mucinous ovarian tumors in the Disease Status TMA

Percentage within each staining category is provided in parentheses. Statistical significance was evaluated using Fisher’s exact test for data in 2 x 2 tables [43]; and the Mehta and Patel version of Fisher’s exact test for data in R x C tables [44]. † p=0.0074 for overall test; p=0.0015 for benign vs. invasive; p>0.05 for benign vs. borderline/LMP; p=0.036 for borderline/LMP vs. invasive. ‡ p=0.0036 for overall test; p=0.012 for benign vs. invasive; p>0.05 for benign vs. borderline/LMP; p=0.014 for borderline/LMP vs. invasive.

Page 6: Legends for supplementary figures Fig s1. A: Western blot showing the suppression of SRp20 expression by Dox induction of SRp20 siRNAs in SKOV3 sublines.

Table s3. Distribution of Evaluable and Total Cases by Tumor Stage, Number of Satisfactory Coresand Histologic Subtype in the Tumor Stage TMA Stained for PTB or SRp20

† Evaluable cases require at least two satisfactory cores with staining for either PTB or SRp20.

Tumor Stage

Satisfactory Cores per Case Evaluable

Cases †TotalCases

Histologic Subtype for Evaluable Cases

0 1 2 3 Endometrioid SerousMixed

Epithelial

PTB staining

Stage I 0 0 12 34 46 46 27 15 4

Stage II 0 3 6 23 29 32 10 19 0

Stage III 0 0 10 47 57 57 26 29 2

Stage IV 1 2 11 26 37 40 13 23 1

Total 1 5 39 130 169 175 76 86 7

SRp20 staining

Stage I 1 4 13 28 41 46 25 12 4

Stage II 1 2 10 19 29 32 10 19 0

Stage III 0 0 12 45 57 57 26 29 2

Stage IV 1 5 12 22 34 40 12 21 1

Total 3 11 47 114 161 175 73 81 7

Page 7: Legends for supplementary figures Fig s1. A: Western blot showing the suppression of SRp20 expression by Dox induction of SRp20 siRNAs in SKOV3 sublines.

Categorized stagePTB † SRp20 ‡

Low High Total Low High Total

Early stage (I+II) 50 (66.7%) 25 (33.3%) 75 42 (60.0%) 28 (40.0%) 70

Advanced stage (III+IV) 45 (47.9%) 49 (52.1%) 94 32 (35.2%) 59 (64.8%) 91

Total 95 74 169 74 87 161

Table s4. Intensity of PTB or SRp20 staining in the Ovarian Cancer Stage TMA

Statistical significance was evaluated using Fisher’s Exact test [43]. Staining intensity was categorized as low (score<2.5) or high (score2.5). † p=0.019 for comparison of early stage vs. advanced stage.‡ p=0.002 for comparison of early stage vs. advanced stage.