RAJARAM, Renuga Devi, et al.

Texte intégral

(1)

Article

Reference

Tubular NOX4 expression decreases in chronic kidney disease but does not modify fibrosis evolution

RAJARAM, Renuga Devi, et al.

Abstract

NADPH oxidase 4 (NOX4) catalyzes the formation of hydrogen peroxide (H2O2). NOX4 is highly expressed in the kidney, but its role in renal injury is unclear and may depend on its specific tissue localization.

RAJARAM, Renuga Devi,

et al

. Tubular NOX4 expression decreases in chronic kidney disease but does not modify fibrosis evolution.

Redox Biology

, 2019, vol. 26, p. 101234

DOI : 10.1016/j.redox.2019.101234 PMID : 31247506

Available at:

http://archive-ouverte.unige.ch/unige:122903

Disclaimer: layout of this document may differ from the published version.

1 / 1

(2)

Supplementary Figure 2: Scheme illustrating the strategy employed for the generation of tubular cell specific Nox4 knock-in mice.

Supplementary Figure 3: Gene expression analysis of Renin and Epo in baseline (A).

Hematoxylin and Eosin staining on the sections of Wt and Nox4KI mice (B). Representative images of active caspase-3 immunostaining on the kidney sections of Wt and Nox4KI mice at baseline (C).

Supplementary Figure 4: Expression analysis of cellular stress (cMyc), angiogenesis (Hif1a, Vegf), pro-inflammatory (Il1b, Il18 and Ccl5) and fibrosis (Tgfb1) genes in the kidney cortical extracts of Wt and Nox4KI mice collected after 3 days (A) and 10 days (B) of UUO. The non- obstructed kidneys were used as controls.

Supplementary Figure 5: Representative images of proliferation marker Ki-67 immunostaining on the kidney sections of Wt and Nox4KI mice collected after 3 days of UUO (A). The non- obstructed kidneys were used as controls. Quantification of Ki-67 immunostaining (B) and apoptosis marker active caspase-3 (C) on the kidney sections of Wt and Nox4KI mice collected after 3 days of UUO.

Supplementary Figure 6: Representative images of macrophage marker F4/80 immunostaining and quantification on the kidney sections of Wt and Nox4KI mice collected after 3 days (A) and 10 days (B) of UUO. The non-obstructed kidneys were used as control.

(3)

Suppl. Figure 1 Rajaram et al

50μm

20μm 20μm

50μm 20μm

50μm 100μm

A

B C

D E

F G

FSGS

DN HTN

(4)

Targeted allele

mNOX4 cDNA

CAG STOP Neor

mNOX4 cDNA

CAG STOP Neor DTA

Conditional knock-in allele after flp mediated recombination

mNOX4 cDNA CAG STOP

Constitutive knock-in allele expression in the tubular cells after Pax8Cre mediated recombination

mNOX4 cDNA CAG

mRosa26 exon Homology arm LoxP site

Frt site PolyA tail

F1

R1

(5)

Suppl.Figure 3 Rajaram et al

Wt Nox4KI 0.0

0.5 1.0 1.5 2.0

Normalized mRNA level

Wt Nox4KI 0.0

1.0 2.0 3.0 4.0

Normalized mRNA level

Wt Nox4KI -2.0

0.0 2.0 4.0 6.0

Normalized mRNA level

Wt Nox4KI 0.0

1.0 2.0 3.0

Normalized mRNA level

ReninEpo

A

Cortex Medulla

50μm

Wt Nox4KI

B Wt Nox4KI

C

50μm

(6)

Wt con Nox4KI

con Wt UUO

Nox4KI UUO 0.0

2.0 4.0 6.0 8.0

NormalizedIl1β Expression

ns **

ns

Wt con Nox4KI

con Wt UUO

Nox4KI UUO 0.0

0.5 1.0 1.5 2.0

NormalizedIl18 Expression

nsns ns

Wt con Nox4

KIcon Wt UUO

Nox4

KIUUO 0.0

5.0 10.0 15.0

NormalizedCcl5 Expression

ns*** ***ns

10d

Wt con Nox4KI

con Wt UUO

Nox4KI UUO 0.0

5.0 10.0 15.0 20.0

NormalizedcMyc Expression

ns** ***ns

Wt con Nox4KI

con Wt UUO

Nox4KI UUO 0.0

0.5 1.0 1.5

NormalizedHif1α Expression

nsns nsns

Wt con Nox4

KIcon Wt UUO

Nox4

KIUUO 0.0

5.0 10.0 15.0

NormalizedIl1β Expression

ns* ** ns

Wt con Nox4

KIcon Wt UUO

Nox4

KIUUO 0.0

0.5 1.0 1.5 2.0 2.5

NormalizedIl18 Expression

nsns nsns

Wt con Nox4

KIcon Wt UUO

Nox4KI UUO 0.0

10.0 20.0 30.0 40.0

NormalizedCcl5 Expression

ns** ***ns Wt con

Nox4KI con

Wt UUO Nox4KI

UUO 0.0

0.5 1.0 1.5 2.0

NormalizedVegf Expression

ns**** ****ns

B

Wt con Nox4

KIcon Wt UUO

Nox4KI UUO 0.0

2.0 4.0 6.0 8.0

NormalizedTgfβ1 Expression

ns**** ***ns

(7)

Suppl. Figure 5 Rajaram et al

A

Wt con Nox4

KIcon Wt UUO

Nox4

KIUUO 0.0

10.0 20.0 30.0

% of Ki-67 +ve cells

ns**** ****ns

B C

100μm

Nox4

KI

- Con Wt- UUO Nox4

KI

- UUO Wt- Con

Wt con Nox4

KIcon Wt UUO

Nox4

KIUUO 0.0

0.2 0.4 0.6 0.8 1.0

% of caspase- 3 +ve cells ns ns** **

(8)

50μm

Nox4KI

50μm

B Con UUO

WtNox4KI

Wt con Nox4

KIcon Wt UUO

Nox4

KIUUO -2.0

0.0 2.0

% of F4/8

Wt con Nox4

KIcon Wt UUO

Nox4

KIUUO 0.0

10.0 20.0 30.0

% of F4/80 +ve area

ns**** ** * 10d

10d

Figure

Updating...

Sujets connexes :