D
(L4L3) region of the L-PK promoter ChREBP-Mlx ChORE L4 L3 -169 -144 -125 DR-1 HNF4
A
Glc R e la ti ve ( L -P K /3 6 B 4 ) m R N A l ev el sB
R el a ti ve ( L -P K /3 6B 4) m R N A le ve ls 0 5 10 15 20 25 30 35Low High Low High
-
+
Glc siRNA FXR-
+
***
**
***
Vehicle GW4064 FXR-siRNA FXR
+
E
**
**
R el at iv e lu ci fe ra se a c ti vi tyGW4064Glc Low
-
High-
High+
Low High Vehicle CDCA
***
***
C
P la sm a g lu c o se ( m g /d l) F RF RF + INT-747 0 1 2 3 4 5 R el a ti ve ( L -P K /2 8S ) m R N A l ev el s***
**
0 50 100 150 200**
**
F RF RF + INT-747 0 5 10 15 20 25 Vehicle GW4064***
*
Low High Vehicle WAY362450***
**
Low High Vehicle INT-747***
**
Low High Glc R e la ti ve ( L -P K /3 6 B 4 ) m R N A l ev el s Actin 1.0 0.6Figure 2. Caron S & Huaman Samanez C
GW4064 WB : ChREBP WB : FXR 1 2 3 4 5- - + - +
95 kDa 59 kDa IP : FXR AB1 AB2B
D
ChREBP Lamin A/C Low-
-High GW4064 Glc 1 2 3 4
+
+
95 kDa 67 kDaA
C
GST GST-FXR ChREBP - - + Mlx HNF4 Input 20% GW4064 FXR fragments F1-105 F106-196 F197-284 F215-300 F301-384 F215-472 Full length FXR 1 105 AF-1 DBD LBD 1 124 196 241 472 106 196 197 284 215 300 301 384 215 472 GST GST-FXR - - + Input 20% GW4064 ChREBP ChREBP ChREBP ChREBP ChREBP ChREBP 1 2 3 4 1 2 3 4R el a ti ve L -P K p ro m o te r o c cu p an c y ChREBP Low High Glc High - -GW4064 +
***
**
R el a ti ve L -P K p ro m o te r o cc u p an c y HNF4 Low High Glc High - -GW4064 +*
R e la ti ve L -P K p ro m o te r o cc u p an c y FXR Low High Glc High - -GW4064 +Figure 4. Caron S & Huaman Samanez C
B
C
E
F
D
GST GST-SMRT FXR - - + 1 2 3 4 Input 10% GW4064A
Low High Low High
Si SMRT R el at iv e (L -P K /3 6B 4 ) m R N A le ve ls - + Glc - + Vehicle GW4064
** **
5 10 15 20 25 30 35 40***
2 4 6 8 10 12 14 R el a ti ve ( L -P K /3 6B 4) m R N A l ev el sLow High Low High
TSA Glc - - + + Vehicle GW4064
***
***
**
SMRT-siRNA SMRT
+
1.0 0.4 R el a ti ve L -P K p ro m o te r o cc u p an c y p300 Low High Glc High - -GW4064 +*
**
R el a ti ve L -P K p ro m o te r o c cu p an cy CBP Low High Glc High - -GW4064 +*
R el at iv e L -P K p ro m o te r o cc u p an cy SMRT Low High Glc High - -GW4064 +*
R el at iv e L -P K p ro m o te r o cc u p an cy H3K9-Ac Low High Glc High - -GW4064 +*
Actin*
A
FAS 0 2 4 6 8 ApoCIII R el at iv e m R N A le ve ls Low High Glc 0.0 0.5 1.0 1.5 2.0 0.0 0.5 1.0 1.5 2.0 2.5 PNPLA3 TxNIP Low High 0 5 10 15 20 Low High Glc Vehicle GW4064 R e la ti ve m R N A le ve lsB
ChREBP R el at iv e T x N IP p ro m o te r o cc u p an c y 0 20 40 60 0 1 2 3 4 5 FXR 0 2 4 6 8 10 SMRT R el at iv e T x N IP p ro m o te r o cc u p an c y 0 2 4 6 8 H3K9-Ac Low High GlcGW4064 Low High High- + + Low High High- + +
0 2 4 6 8 10 12 14 p300 R e la ti ve T xN IP p ro m o te r o cc u p a n cy 0 1 2 3 4 5 6 CBP Glc
GW4064 Low High High- + + Low High High- + +
Glc
GW4064 Low High High- + + Low High High- + +
*
*
***
*
Figure 6. Caron S & Huaman Samanez C
0.0 0.5 1.0 1.5 2.0 2.5 Low High Glc 9 0 1 2 3 4 5 6 7 8 Low High Glc 0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 Low High Glc 0.0 0.5 1.0 1.5 2.0 2.5 Low High Glc 0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 Low High Glc 0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 Low High Glc PGM1 HK3 TPI1 PGAM1 ACSS1 LDHA R el a ti ve m R N A le v el s R e la ti ve m R N A le ve ls R e la ti ve m R N A le ve ls Vehicle GW4064**
*
**
*
**
*
*
*
*
*
*** *
L-PK promoter ChREBP ChORE L4 L3 -169 -144 -125 DR1 HNF4 p300 FXR CBP
High glucose
L-PK promoter ChREBP ChORE L4 L3 -169 -144 -125 DR1 HNF4 p300 CBP FXRHigh glucose + FXR activation
SMRT
