Correlation between ANME-2b and Seep-1g
fluorescent signal
Correlation between Seep-1g and nifH
fluorescent signal
Correlation between ANME-2b and nifH
fluorescent signal
Pearson’s correlation coefficient,
PC=0.661
,
p-value-100%, Costes’ method
Pearson’s correlation coefficient,
PC=0.893
,
p-value=100%, Costes method
Pearson’s correlation coefficient,
PC=0.837
,
p-value =Costes’ method
Manders’ correlation coefficients, after
thresholding,
M1=0.859, M2=0.552
Manders’ correlation coefficients, after
thresholding,
M1=0.964, M2=0.733
Manders’ correlation coefficients, after
thresholding,
M1=0.805, M2=0.905
a. Scatterplots of pixel intensities
P
C
=0.66
1
,
P
C
=0.89
3
,
P
C
=0.837
,
ANME-2b
Seep-1g
nifH
DAPI
Supplementary Figure 9.
Colocalization analysis of HCR-FISH experimental data to investigate nitrogenase expression by nifH targeted probes.
A
NME-2b is stained in the cy3 channel, Seep-1g in the FITC channel and the B1 amplifiers binding nifH initiator probes are visua
lized in the cy5
channel a. Scatterplots of pixel intensities of the FITC, cy3 and cy5 channel suggest that there is correlation between the See
p-1g signal and
nifH signal, as well as correlation between ANME-2b and the nifH signal. However, there appears to be more noise in the latter,
rather than just
a linear correlation between the ANME-2b and nifH signals. An equally high PC between between Seep-1g and nifH, and ANME-2b and
nifH is
suggestive of nifH expression in ANME-2b as well. This is not an observation that is visually obvious and the lower intensity s
ignal could come from nifH
probes designed to target Seep-1g, binding ANME-2b nifH with lower efficiency. The Manders’ coefficients suggest that almost al
l of the Seep-1g
cells correlate with nifH signal while the same is not true of the colocalization of nifH signal with ANME-2b.