Skip to main content
. Author manuscript; available in PMC: 2021 Jul 28.
Published in final edited form as: Nat Microbiol. 2021 Jan 28;6(4):455–466. doi: 10.1038/s41564-020-00850-3

Extended Data Fig. 2. Asian ZIKV infection, but not African ZIKV elicits in vitro calcification in pericytes.

Extended Data Fig. 2

Human fetal pericytes were infected with different ZIKV strains at MOI of 0.5 or mock (PBS) as control (n=4 biologically independent cells per group). (a) At 3 dpi, cell viability was quantified. See Supplementary Data Fig. 1 for gating strategy. (b) BMP4, BMP6, BMP7, BMP9 and NOG expression between ZIKV MR766- and ZIKV H/PF/2013-infected pericytes at 1, 3 and 4 dpi (n=6 biologically independent cells per group). Data in a and b are presented as mean ± SEM in box plots showing the upper (75%) and lower (25%) quartiles, with the horizontal line as the median and the whiskers as the maximum and minimum values observed. (c) Osteogenic gene expression between ZIKV MR766- and ZIKV IbH30656-infected pericytes was normalized to GAPDH and expressed as fold change relative to mock controls (n=3 biologically independent cells per group). Data are presented as mean ± SEM. (d) At 8 and 14 dpi, mock- or ZIKV-infected human fetal pericytes were subjected to Alizarin red staining for calcium deposition. Data are representative of two independent experiments. (e-f) Alizarin red staining were performed on mock- or ZIKV-infected SK-N-SH and U251 at 14 and 21 dpi. Data are representative of four independent experiments. (g) Human primary fetal pericytes (n=5) and astrocytes (n=6) were infected with PBS (mock) or ZIKV PRVABC59 at MOI of 0.5. At 3 dpi, osteogenic gene expressions were normalized to GAPDH and expressed as fold changes relative to mock controls. Data are presented as mean ± SEM and are representative of two independent experiments. (h) RNAseq data generated from Nature Neuroscience; 2017; 20(9); p1209–1212, doi: 10.1038/nn.4612 (GEO ID: GSE87750) was reanalyzed for osteogenic gene expression. Human primary peripheral neurons were infected with ZIKV PRVABC59 of MOI 0.4 and harvested at 3 dpi. Statistical analyses were performed using one-way ANOVA with Tukey’s posttest (a and c). ***P< 0.001 and ****P< 0.0001. Exact P values in a compared between ZIKV MR766 and ZIKV H/PF/2013 group (P= 0.0008), ZIKV MR766 and ZIKV PRVABC59 group (P= 0.0005), ZIKV IBH30656 and ZIKV H/PF/2013 group (P= <0.0001) and ZIKV IBH30656 and ZIKV PRVABC59 group (P= <0.0001).