Additional file 1 of Systemic administration of choline acetyltransferase decreases blood pressure in murine hypertension
Creators: name: Stiegler, Andrew
name: Li, Jian-Hua
name: Shah, Vivek
name: Tsaava, Tea
name: Tynan, Aisling
name: Yang, Huan
name: Tamari, Yehuda
name: Brines, Michael
name: Tracey, Kevin J.
name: Chavan, Sangeeta S.
Publisher: name: figshare
Year: 2021
Subjects: subject: Biochemistry
subject: Medicine
subject: Genetics
subject: FOS: Biological sciences
subject: Molecular Biology
subject: Neuroscience
subject: Physiology
subject: Pharmacology
subject: Biotechnology
subject: Cancer
subject: Mental Health
subject: Hematology

description: Additional file 1: Figure S1. ChAT administration decreases SBP and DBP. A Change in normalized SBP in the 3 h period after ChAT administration. SBP is normalized to the 30-min period before ChAT or vehicle administration. After normalization, area under the curve is calculated from a baseline of 100%. Negative values correspond to decrease in SBP. n = 7–11. *p &lt; 0.05, one-way ANOVA with Dunnett’s correction. B Change in normalized DBP in the 3 h period after ChAT administration. DBP is normalized to the 30-min period before ChAT or vehicle administration. After normalization, area under the curve is calculated from a baseline of 100%. Negative values correspond to decrease in DBP. Data are represented as individual mouse data points, mean ± SEM. n = 7–11. ns not significant one-way ANOVA with Dunnett’s correction. Figure S2. ChAT administration increases heart rate. Change in heart rate after ChAT administration. Heart rate is averaged over the 30 min pre-administration. Change is calculated as the pre-administration baseline subtracted from the average heart rate in the 3 h post-administration period. Data are represented as individual mouse data points, mean ± SEM. n = 5–11, *p &lt; 0.05, t-test. Figure S3. ChAT administration does not affect body temperature. Change in body temperature after ChAT administration. Body temperature is averaged over the 30 min pre-administration. Change is calculated as the pre-administration baseline subtracted from the average body temperature in the 3 h post-administration period. Data are represented as individual mouse data points, mean ± SEM. n = 5–11, ns not significant, t-test. Figure S4. ChAT administration increases activity levels. Change in activity after ChAT administration. Activity is averaged over the 30 min pre-administration. Change is calculated as the pre-administration baseline subtracted from the average activity in the 3 h post-administration period. Data are represented as individual mouse data points, mean ± SEM. Vehicle n = 5–11, ns not significant, t-test. Figure S5. PEGylation increases molecular weight without decreasing specific activity. A SDS-PAGE of purified recombinant human ChAT with and without PEGylation. With PEGylation, molecular weight increases by approximately 20 kDa. B comparison of specific activity of ChAT and PEG-ChAT, as determined by colorimetric activity assay. Data represented as one assay performed in triplicate. ****p &lt; 0.0001, one-way ANOVA with Dunnett’s correction. Figure S6. PEG-ChAT administration decreases both SBP and DBP. A change in normalized SBP in the 3 h period after PEG-ChAT administration. SBP is normalized to the 30-min period before PEG-ChAT (0.1 or 1 mg/kg) or vehicle administration, and area under the curve is calculated from a baseline of 100%. Negative values correspond to decrease in SBP. Data are represented as individual mouse data points, mean ± SEM. n = 7–11 per group. ***p &lt; 0.001, one-way ANOVA with Dunnett’s correction. B change in normalized DBP in the 3 h period after PEG-ChAT administration. SBP is normalized to the 30-min period before PEG-ChAT (0.1 or 1 mg/kg) or vehicle administration, and area under the curve is calculated from a baseline of 100%. Negative values correspond to decrease in DBP. Data are represented as individual mouse data points, mean ± SEM. n = 7–11 per group. **p &lt; 0.01, one-way ANOVA with Dunnett’s correction. Figure S7. PEG-ChAT administration does not induce long-term changes in heart rate, body temperature, or activity levels. A HR, B temperature and C activity in the 24 h period post PEG-ChAT administration. Gray background indicates dark cycle. Data are averaged every hour and represented with lines as group mean ± SEM. ns not significant, two-way ANOVA with Sidak’s multiple comparisons test. Figure S8. PEG-ChAT administration during dark cycle induces decreases in SBP and DBP in the following light cycle. A mean SBP, B mean DBP in light cycle. PEG-ChAT was administered at the middle of the dark cycle (6 h), and MAP is averaged 6–18 h post-PEG-ChAT administration. Data are represented as individual mouse data points, mean ± SEM. Vehicle: n = 12, PEG-ChAT 0.1 mg/kg: n = 20, PEG-ChAT 1 mg/kg: n = 21, *p &lt; 0.05, ***p &lt; 0.001, ****p &lt; 0.0001, one-way ANOVA with Dunnett’s correction.