Human and Mouse Neuroinflammation Markers in Niemann-Pick Disease, type C1
Stephanie M. Cologna1*, Celine V.M. Cluzeau1*, Nicole M. Yanjanin1, Paul S. Blank2, Michelle
K. Dail1, Stephan Siebel1, Cynthia L. Toth1, Christopher A. Wassif1, Andrew P. Lieberman3 and
Forbes D. Porter1#
1
Program in Developmental Endocrinology and Genetics, Section on Molecular Dysmorphology
NICHD, NIH, DHHS, Bethesda, MD, USA 20892
2
Program in Physical Biology, Section on Membrane & Cellular Biophysics, NICHD, NIH,
DHHS, Bethesda, MD, USA 20892
3
Department of Pathology, University of Michigan, Ann Arbor, MI USA, 48109
* Indicates equal contributions by these authors
#Corresponding Author
Forbes D. Porter, M.D., Ph.D.
PDEGEN, NICHD, NIH, DHHS
Bld. 10, Rm. 9D42
10 Center Dr.
Bethesda, MD 20892
Phone: 301-435-4432
Fax: 301-480-5791
Email: fdporter@mail.nih.gov
Supplemental Figure 1: Validation of mouse inflammation PCR array results in the 7week-old cerebral cortex tissue. In order to validate the results obtained on the PCR array, we
used qPCR to determine the expression pattern of 5 genes in 7-week-old Npc1+/+ (white
columns) and Npc1-/- (black columns) cerebral cortex. Control mice were used as reference
group for the relative quantitation. Mean and standard deviation are shown for each group (N=4).
An unpaired t-test with Welch’s correction, when necessary, was performed to determine the
significance of the difference in means between the log10 values of control and mutant mice
relative quantitation at each age.
Supplemental Figure 2: Weight loss and survival of Npc1 mice with Ccl3 deficiency. Mean
weight measurement for female (a) and male (b) mice of the different genotypes. Numbers of
animals in each group are indicated below each graph. (c) Kaplan-Meier survival curves of mice
of the different genotypes. Age at death was approximated by their age when sacrificed,
corresponding to a 20% weight loss.
Supplemental Figure 3: Validation of human inflammation PCR array results. We used
qPCR to determine the expression pattern of 10 genes in control (open circles) and NPC1
patients (black squares) frontal cortex and cerebellum to validate the results obtained on the PCR
array. Control group was used as reference for the relative quantitation. Mean and standard
deviation are shown for each group (N=4 for cerebellum; N=7 for frontal cortex). An unpaired ttest with Welch’s correction, when necessary, was performed to determine the significance of the
difference in means between the log10 values of control and patient relative quantitation.
Supplemental Figure 4: Cerebrospinal fluid interleukins without altered expression in
NPC1 patients. Log10 transformed concentrations of 6 interleukins in cerebrospinal fluid from
controls (circles) and NPC1 patients (squares). Mean and standard deviation are shown for each
group. An unpaired t-test with Welch’s correction when necessary was performed to determine
the significance of the difference in means between controls and patients.
Supplemental Figure 5: Cerebrospinal fluid inflammation molecules without altered
expression in NPC1 patients. Log10 transformed concentrations of 4 molecules in
cerebrospinal fluid from controls (circles) and NPC1 patients (squares). Mean and standard
deviation are shown for each group. An unpaired t-test with Welch’s correction when necessary
was performed to determine the significance of the difference in means between controls and
patients.
Supplemental Figure 6: Comparison of CSF levels of inflammation markers with respect to
miglustat treatment. (a, c, e) The group of NPC1 patients was separated between patients
receiving miglustat treatment (triangles) and the ones not receiving it (inverted triangles). Mean
and standard deviation are shown for each group. An unpaired t-test with Welch’s correction
when necessary was performed to determine the significance of the difference in means between
controls and patients. (b, d, f) Percent change in each inflammatory marker CSF concentration
over time after initiation of miglustat treatment for 5 patients.