Figure 4.

Naturally Occurring TMEM41B SNPs Negatively Impact Flavivirus Infection but Can Maintain Normal Lipid Distribution in Cells

(A) Table shows the frequency of a SNP (rs78813996) in TMEM41B in several human populations.

(B) WT, TMEM41B KO, and TMEM41B KO HAP1 cells expressing WT or TMEM41B SNP variants infected with YFV 17D. Cells were analyzed by flow cytometry and plotted as a percentage of viral antigen-positive cells.

(C) Infected as in (B), and supernatants were collected and titrated by tissue culture infectious dose (TCID₅₀/mL) assay on Huh-7.5 cells. The two-tailed statistical Student’s t test was used to determine statistical significance and is depicted with an asterisk indicating a p value < 0.05.

(D) Same as (B) infected with ZIKV, DENV-GFP, WNV-GFP, and hPIV-3-GFP.

(E) WT and TMEM41B KO Huh-7.5 cell clones stained with Nile red to visualize lipid droplets.

(F) Cumulative frequency of droplets plotted versus droplet area (μm²) for six independent single-cell clones generated with two independent sgRNAs. Inset shows the mean lipid droplet area (μm²) for the six TMEM41B KO clones compared to WT Huh-7.5 cells.

(G) Mean lipid droplet area (μm²) for WT, TMEM41B KO, and TMEM41B KO Huh-7.5 cells (clone 1-1) expressing the indicated TMEM41B variants. Cells were analyzed by flow cytometry and plotted as a percentage of viral antigen positive cells. Error bars depict SD for n = 3 replicates.