supplementary figure 1

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Supplementary Figure 1 A B C D Relative light units (RLU) 0 20000 40000 60000 80000 100000 120000 Line I Line 2 + - - + Dox *** *** T7-PELP1 T7-PELP1 WT MMTV-rtTA TetO-PELP1 MMTV-rtTA TET0-PELP1 Line 1 MMTV-rtTA TET0-PELP1 Line 2 rtTA rtTA + Dox Figure S1. Characterization of PELP1 Tg mice . A, Schematic representation of bitransgenic mouse model system. PELP1 transgene construct consists of a full-length T7 and His-tagged human PELP1 cDNA linked to luciferase gene reporter through an internal ribosomal entry site (IRES). B, pTetO-PELP1 mice crossed with mammary gland–specific rtTA mice (MMTVrtTA) to establish two independent MMTVrtTA-TetO-PELP1 transgenic lines. Identification and establishment of the transgenic lines was done by using PCR analysis with transgene-specific primers. C, To monitor luciferase activity in vitro, mammary gland homogenates of aged-matched experimental nulliparous adult female bitransgenic mice treated with or without doxycycline were assessed by using a Dual Luciferase Assay System. All experimental data points were generated from three biological replicates per transgenic line. Statistical significance was determined by using Student’s t test.****p<0.0001***p<0.001; ; *p<0.05. D, Human PELP1 transcript induction in mammary tissue of 6- month-old virgin mice by real-time RT-PCR analysis. Line 1 and Line 2 displayed two- and five-fold increases, respectively. Statistical Relative expression (norm alized to actin) 0 2 4 6 8 * * con Line Line 2

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Supplementary Figure 1. ***. 120000. 100000. ***. 80000. 60000. Relative light units (RLU). 40000. 20000. 0. Dox. +. -. +. -. Line I. Line 2. rtTA. rtTA. A. MMTV- rtTA TET0-PELP1 Line 1. MMTV- rtTA TET0-PELP1 Line 2. B. TetO-PELP1. MMTV- rtTA. WT. + Dox. T7-PELP1. - PowerPoint PPT Presentation

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Page 1: Supplementary Figure 1

Supplementary Figure 1

A B

C D

Rel

ativ

e lig

ht u

nits

(R

LU)

0

20000

40000

60000

80000

100000

120000

Line I Line 2

+ - -+Dox

***

***

T7-PELP1

T7-PELP1

WT

MM

TV

-rtT

A

TetO

-PE

LP1

MM

TV

-rtT

AT

ET

0-P

ELP

1Li

ne 1

MM

TV

-rtT

AT

ET

0-P

ELP

1Li

ne 2rtTA

rtTA

+ Dox

Rel

ativ

e ex

pres

sion

(nor

mal

ized

to a

ctin

)

0

2

4

6

8

*

*

con Line Line 2

Figure S1. Characterization of PELP1 Tg mice . A, Schematic representation of bitransgenic mouse model system. PELP1 transgene construct consists of a full-length T7 and His-tagged human PELP1 cDNA linked to luciferase gene reporter through an internal ribosomal entry site (IRES). B, pTetO-PELP1 mice crossed with mammary gland–specific rtTA mice (MMTVrtTA) to establish two independent MMTVrtTA-TetO-PELP1 transgenic lines. Identification and establishment of the transgenic lines was done by using PCR analysis with transgene-specific primers. C, To monitor luciferase activity in vitro, mammary gland homogenates of aged-matched experimental nulliparous adult female bitransgenic mice treated with or without doxycycline were assessed by using a Dual Luciferase Assay System. All experimental data points were generated from three biological replicates per transgenic line. Statistical significance was determined by using Student’s t test.****p<0.0001***p<0.001; ; *p<0.05. D, Human PELP1 transcript induction in mammary tissue of 6-month-old virgin mice by real-time RT-PCR analysis. Line 1 and Line 2 displayed two- and five-fold increases, respectively. Statistical significance was determined by using Student’s t test; n=3 per group, *p<0.05.