Figure1

A mpeg1:mCherry 11 dpf larvae B Macrophage C TUNEL+

DMSO Prednisolone number in larvae macrophages
**

Macrophages/section
**

% of macrophages
80
8 60
40
4
20
0 0
DMSO Pred. DMSO Pred.
D adult mpeg1:mCherry head sections E Macrophages in F Macrophages in
Brightfield mpeg1:mCh Overlay adult heads adult fins

Macrophages/mm tissue
20 20 **

Macrophages/mm fin
*
DMSO

15 15
10 10
5 5
Pred.

0 0
DMSO Pred. DMSO Pred.

G Amount of myelomonocytes H Amount of lymphocytes I qRT-PCR on fins

in the kidney of adult fish in the kidney of adult fish 160

cxcl8a expression (%)

Myelomonocytes (%

15 * 15 *
120
Lymphocytes (%
of all cells)

of all cells)

10 10
80
5 5 40

0 0 0
DMSO Pred. DMSO Pred. DMSO Pred.

J qRT-PCR on fins K Neutrophils in

150 the regenerate
0,2
tnf-a expression (%)

mpx:GFP + cells/µm2

100
0,1
50

0 0
DMSO Pred. DMSO Pred.
 Figure 2

A Alizarin red staining B Alizarin red stained C Brightfield Calcein staining

bone in larvae

DMSO
DMSO
**
140

% Mineralized area
120
100
80
60
40

Pred.
Pred.

20
0
DMSO Pred.

D Calcein stained E Adult uninjured fins F Alizarin red staining

bone in larvae DMSO Prednisolone of the uninjured fin
120 ** 40 *

100

Unstained region of
Alizarin red staining

30
% Calcified area

80

the fin (%)

60 20
40
10
20
0 0
DMSO Pred. DMSO Pred.

G H
Vertebral bone volume Length of the regenerate after caudal fin amputation
0.040 *** ***
0.035 3 ***
***

0.030
BV (mm3)

Length (mm)

***
0.025 DMSO
2
0.020 * Pred.
0.015
1
0.010
0.005
0 0
DMSO Pred. 4 7 9 14 17 22 27 dpa
GFP

GFP

J Osterix:nGFP K Alizarin red staining

BF

BF

I
regenerate length at 14 dpa of regenerating fins

3
Length of alizarin red+
osterix:nGFP 14 dpa

4 **
Prednisolone

Length (mm)

bone (mm)

3 2
DMSO

2
1
1

0 0
DMSO Pred. DMSO Pred.

L Brighfield GFP M 7 dpi alizarin red stained N Regenerated skull

skull injuries bone area
DMSO

120
osterix:nGFP heads

DMSO Prednisolone
Filled area (%)

80

40
Pred.

0
DMSO Pred.
 Figure 3

A osterix:nGFP intensity in B osterix:nGFP C TUNEL+ osteoblasts

regenerates intensity in skull injuries 4 dpa
50 0,008
Intensity (arbitrary units)

**

Intensity (arbitrary units)

3
40 0,006

% of osteoblasts
2 DMSO 30
Pred. 0,004
1 20
0,002
0 10
0 100 200 300 0,000
0
Distance from amputation plane (µm) DMSO Pred. DMSO Pred.

D PCNA+ osteoblasts 4 dpa E Pred. DMSO untr. Brightfield PCNA osterix:nGFP Overlay + DAPI
**
PCNA+ OB/mm fin

15
4 dpa treated fins

*
10

0
untr. DMSO Pred.

F qRT-PCR on G 0 hpa 3 dpa H qRT-PCR on I DMSO Pred.

2 dpa regenerates BF GFP BF GFP 4 dpa regenerates BF GFP BF GFP
runx2a expression (%)

osteocalcin:GFP 10 dpa
bglap expression (%)

120
DMSO

120
runx2:GFP

80 80

40 40
Pred.

0 0
DMSO Pred. DMSO Pred.
 Figure 4

A TRAP staining in larvae B TRAP staining

in larvae
DMSO 80 **
60

TRAP+ cells
40
20
Pred.

0
DMSO Pred.

C DMSO Pred. D TRAP+ cells in E TRAP+ cells in

the regenerate the stump
150 *** 80
TRAP staining in
regenerating fins

60

TRAP+ cells
TRAP+ cells

100
40
50 20

0 0
DMSO Pred. DMSO Pred.