Disease Models & Mechanisms is committed to maintaining the integrity … image... ·...
Transcript of Disease Models & Mechanisms is committed to maintaining the integrity … image... ·...
Best Practices: gels, blots and images in DMM figures
Disease Models & Mechanisms is committed to maintaining the integrity of the scientific record. We therefore screen all digital images in accepted manuscripts for signs of inappropriate manipulation.
Any article that does not meet the following requirements will be delayed in the publication process; your paper may be rejected on the basis of non-compliance with these guidelines.
Do not add to, alter, enhance, obscure, move or remove a specific feature of an image – the focus should be on the data rather than its presentation (e.g. do not ‘clean up’ backgrounds or remove/obscure imperfections and non-specific bands)
Adjustments should be applied to the whole image so no specific feature of the original data, including background, is obscured, eliminated or misrepresented as a consequence. Any alterations, such as non-linear adjustments (e.g. changes to gamma settings), must be disclosed in the appropriate figure legends and in the Materials and Methods section.
e.g. Original data Extra bands added data Extra bands detected
Non-linear adjustment Non-linear adjustment detected
The splicing of multiple images to suggest they come from a single micrograph or gel is not allowed.
Any grouping or consolidation of data (e.g. removal of lanes from gels and blots or cropping of images) must be made apparent (i.e. with dividing lines or white spaces) and should be explicitly indicated in the figure legends.
e.g. Submitted figure Corrected figure with splices marked
At least several band widths should be retained above and below cropped bands
A positive and a negative control and a set of molecular weight markers must be indicated on all images of gels and blots. High-contrast gels and blots are unacceptable (i.e. no white backgrounds) – grey backgrounds are expected unless otherwise justified
Not acceptable Acceptable
The same data in whole or part should not be presented in multiple figures (e.g. loading controls; different exposures of the same gel), unless explicitly stated and justified
Previously published data in whole or in part (e.g. loading controls) should not be presented
Better �gure layout can help reviewers and readers to appreciate your science
DMM text is spread over two columns (grey boxes) into which �gures �t (yellow boxes)
Legend Legend
Legend
Legend
Legend
1. Consider �gure layout in the context of single or double columns (the yellow boxes above).
2. Within the �gure (yellow box) maximise information (data) and minimise background.
3. Consider that di�erent data work on di�erent scales. Images of cells and western blots shouldbe large enough to see the relevant features. Information in graphs can often be read when smaller (see page 2).
ITSN-1
β-Actin
Control
1-1 1-2
LS
188
49
A
Ex. v
irus
(B5)
/ Ac
tinA
ctin
Control ITSN1 1 ITSN1 2B
0
25
50
75
100
125
No.
act
in ta
ils p
er c
ell
0
50
75
100
125
25
Control
1-1 1-2
******
0
10
20
30
Control
1-1 1-2
% e
x-vi
rus
indu
cing
act
in
0
10
20
30***
***
C
Single column / half-page width
- Data �ts nicely in area- Background minimised- Western and graph size ok- Cell images too small
Ex. v
irus
(B5)
/ A
ctin
Act
in
Control ITSN1 1 ITSN1 2
0
25
50
75
100
125
No.
act
in ta
ils p
er c
ell
0
50
75
100
125
25
Control
1-1 1-2
******
0
10
20
30
Control
1-1 1-2
% e
x-vi
rus
indu
cing
act
in
0
10
20
30***
***
ITSN-1
β-Actin
Control
1-1 1-2
LS
188
49
A B
C
Double column / full-page width
- Data �ts nicely in area - Background minimised- Relevant features in images and graphs are visible
Ex. v
irus
(B5)
/ Ac
tinAc
tin
Control ITSN1 1 ITSN1 2
ITSN-1
β-Actin
Control
1-1 1-2
LS
188
49
A B
C
0
25
50
75
100
125
No.
act
in ta
ils p
er c
ell
0
50
75
100
125
25
Control
1-1 1-2
******
0
10
20
30
Control
1-1 1-2
% e
x-vi
rus
indu
cing
act
in
0
10
20
30***
***
1.5 columns / box format
- Data layout not optimal - Background content not minimised- Western blot and cell images too small- Graphs bigger than they need to be
Below we illustrate points 1 to 3 using data from Fig. 5, Humphries et al., Journal of Cell Science (2014), 127, 673–685.
See page two for more layout examples and tips
We prefer single or double column width layouts
0 2 4 6 8 10 12 14 16 18 20 22 240.0
0.2
0.4
0.6
0.8
1.0
Time (seconds)N
orm
alis
e in
tens
ity
0 2 4 6 8 10 12 14 16 18 20 22 240
0.2
0.4
0.6
0.8
1.0
WTH208D
Percentage recovery Half-life (s)
85.1±0.0187.4±0.01
2.72±0.201.18±0.10
ΔNckH208D
88.7±0.01 0.86±0.06
Time (s)
A
Plaq
ue s
ize
(mm
)
0.0
0.5
1.0
1.5
2.0
-/- WT
H208D
H208DΔNck
GFP-N-WASP
CB
N-W
ASP
-ΔN
ck-H
208D
ActinGFP Ex. virus (B5)
N-W
ASP
-ΔN
ck
n.s.
-/-
WT
H208D ΔNck-H208D
GFP-N
-WA
SP
0
25
50
75
100
0
25
50
75
100
% c
ells
with
act
in ta
ils
0
25
50
75
100
WT
H208D
No.
act
in ta
ils p
er c
ell
0
50
75
100
25
ΔNckΔNck
n.s. **
H208D WT
H208DΔNck
ΔNck
H208D
**
0
20
40
60
*** **n.s.
0
40
60
20
WT
H208DΔNck
ΔNck
H208D
% e
x-vi
rus
with
act
in ta
il
D E
0
25
50
75
100
0
25
50
75
100
% c
ells
with
act
in ta
ils
0
25
50
75
100n.s.
WT
H208D
**
∆Nck
H208D0
25
50
75
100**
WT
H208D∆Nck
H208D
Num
ber o
f act
in ta
ils p
er c
ell
**
A36-Y132F
ActinGFP Ex. virus (exB5)
N-W
ASP
N-W
ASP
- H20
8DN
-WA
SP-Δ
Nck
-H20
8D
A36-Y132F
0 2 4 6 8 10 12 14 16 18 20 22 240.0
0.2
0.4
0.6
0.8
1.0
Time (seconds)
Nor
mal
ise
inte
nsity
0 2 4 6 8 10 12 14 16 18 20 22 240
0.2
0.4
0.6
0.8
1.0
WTH208D
Percentage recovery Half-life (s)
85.1±0.0187.4±0.01
2.72±0.201.18±0.10
ΔNckH208D
88.7±0.01 0.86±0.06
Time (s)
A
Plaq
ue s
ize
(mm
)
0.0
0.5
1.0
1.5
2.0
-/- WT
H208D
H208DΔNck
GFP-N-WASP
C
B
N-W
ASP
-ΔN
ck-H
208D
ActinGFP Ex. virus (B5)
N-W
ASP
-ΔN
ck
n.s.
-/-
WT
H208D ΔNck-H208D
GFP-N
-WA
SP
0
25
50
75
100
0
25
50
75
100
% c
ells
with
act
in ta
ils
0
25
50
75
100
WT
H208D
No.
act
in ta
ils p
er c
ell
0
50
75
100
25
ΔNckΔNck
n.s. **
H208D WT
H208DΔNck
ΔNck
H208D
**
0
20
40
60
*** **n.s.
0
40
60
20
WT
H208DΔNck
ΔNck
H208D
% e
x-vi
rus
with
act
in ta
il
D E
0
25
50
75
100
0
25
50
75
100
% c
ells
with
act
in ta
ils
0
25
50
75
100n.s.
WT
H208D
**
∆Nck
H208D0
25
50
75
100**
WT
H208D∆Nck
H208D
Num
ber o
f act
in ta
ils p
er c
ell
**
A36-Y132F
ActinGFP Ex. virus (exB5)
N-W
ASP
N-W
ASP
- H20
8DN
-WA
SP-Δ
Nck
-H20
8D
A36-Y132F
Small images, overly big graphs and white gaps between rows. Images too big, graphs too small and odd white gaps.
Odd ordering , large white gaps and ine�ective use of space. Layout as published maximises information in available space.
0 2 4 6 8 10 12 14 16 18 20 22 240.0
0.2
0.4
0.6
0.8
1.0
Time (seconds)
Nor
mal
ise
inte
nsity
0 2 4 6 8 10 12 14 16 18 20 22 240
0.2
0.4
0.6
0.8
1.0
WTH208D
Percentage recovery Half-life (s)
85.1±0.0187.4±0.01
2.72±0.201.18±0.10
ΔNckH208D
88.7±0.01 0.86±0.06
Time (s)
APl
aque
siz
e (m
m)
0.0
0.5
1.0
1.5
2.0
-/- WT
H208D
H208DΔNck
GFP-N-WASP
C
B
N-W
ASP
-ΔN
ck-H
208D
ActinGFP Ex. virus (B5)
N-W
ASP
-ΔN
ck
n.s.
-/-
WT
H208D ΔNck-H208D
GFP-N
-WA
SP
0
25
50
75
100
0
25
50
75
100
% c
ells
with
act
in ta
ils
0
25
50
75
100
WT
H208D
No.
act
in ta
ils p
er c
ell
0
50
75
100
25
ΔNckΔNck
n.s. **
H208D WT
H208DΔNck
ΔNck
H208D
**
0
20
40
60
*** **n.s.
0
40
60
20
WT
H208DΔNck
ΔNck
H208D
% e
x-vi
rus
with
act
in ta
il
D
E
0
25
50
75
100
0
25
50
75
100
% c
ells
with
act
in ta
ils
0
25
50
75
100n.s.
WT
H208D
**
∆Nck
H208D0
25
50
75
100**
WT
H208D∆Nck
H208D
Num
ber o
f act
in ta
ils p
er c
ell
**
A36-Y132FActinGFP Ex. virus (exB5)
N-W
ASP
N-W
ASP
- H20
8DN
-WA
SP-Δ
Nck
-H20
8D
A36-Y132F
Data are from Fig. 2, Humphries et al., Journal of Cell Science (2014), 127, 673–685.
0 2 4 6 8 10 12 14 16 18 20 22 240.0
0.2
0.4
0.6
0.8
1.0
Time (seconds)
Nor
mal
ise
inte
nsity
0 2 4 6 8 10 12 14 16 18 20 22 240
0.2
0.4
0.6
0.8
1.0
WTH208D
Percentage recovery Half-life (s)
85.1±0.0187.4±0.01
2.72±0.201.18±0.10
ΔNckH208D
88.7±0.01 0.86±0.06
Time (s)
A
Plaq
ue s
ize
(mm
)
0.0
0.5
1.0
1.5
2.0
-/- WT
H208D
H208DΔNck
GFP-N-WASP
CB
N-W
ASP
-ΔN
ck-H
208D
ActinGFP Ex. virus (B5)
N-W
ASP
-ΔN
ck
n.s.
-/-
WT
H208D ΔNck-H208D
GFP-N
-WA
SP
0
25
50
75
100
0
25
50
75
100
% c
ells
with
act
in ta
ils
0
25
50
75
100
WT
H208D
No.
act
in ta
ils p
er c
ell
0
50
75
100
25
ΔNckΔNck
n.s. **
H208D WT
H208DΔNck
ΔNck
H208D
**
0
20
40
60
*** **n.s.
0
40
60
20
WT
H208DΔNck
ΔNck
H208D
% e
x-vi
rus
with
act
in ta
il
D E
0
25
50
75
100
0
25
50
75
100
% c
ells
with
act
in ta
ils
0
25
50
75
100n.s.
WT
H208D
**
∆Nck
H208D0
25
50
75
100**
WT
H208D∆Nck
H208D
Num
ber o
f act
in ta
ils p
er c
ell
**
A36-Y132F
ActinGFP Ex. virus (exB5)
N-W
ASP
N-W
ASP
- H20
8DN
-WA
SP-Δ
Nck
-H20
8D
A36-Y132F