Top scoring HDFs are components of the autophagy pathway.

MERS-CoV (A) and HCoV-229E (B) titers upon KO of top scoring HDFs are displayed in Log 10 TCID 50 /ml. Raw data can be found in Supporting information S1 Data , tabs 6 and 7. MERS-CoV (C) , HCoV-229E (D) , SARS-CoV (E) , and SARS-CoV-2 (F) titers upon reconstitution of TMEM41B, FKBP8, and MINAR1 in respective KO cell lines. Titers are shown relative to WT cell line. Results are displayed as a mean of 3 independent experiments with SD represented by error bars. Raw data can be found in Supporting information S1 Data , tab 8. (G) Quantification of western blot analysis of FKBP8 and TMEM41B in Huh7 WT, KO, and reconstituted cells relative to beta actin. Quantification of MINAR-GFP utilizing an anti-GFP antibody in WT, KO, and MINAR1-GFP reconstituted cells relative to beta actin. Raw western blot images can be found in Supporting information S1 Raw Images. Quantification data can be found in Supporting information S1 Data , tab 9. (H) Immunofluorescence analysis of VSVΔG-CoV-spike-GFP and VSVΔG complemented with G in respective CTRL and KO cells. Scale bar is 40 μm. GFP is depicted in green, and DAPI is shown in blue. A total of 20 to 26 images per condition in 3 independent experiments were acquired using an Evos FL Auto 2 imaging system with a 10× air objective, analyzed and quantified in Fiji. Representative images of 1 out of 3 independent replications are shown. (I) Quantification of CoV-mediated entry visualized in (H). Raw data can be found in Supporting information S1 Data , tab 10. In A and B, statistical analysis was determined by ordinary 1-way ANOVA, Dunnett multiple comparison test, using Nev 2020 version 9.0. In C–G, statistical significance was determined in R (version 4.0.2) using a 2-tailed paired t test to compare titers between each KO and KO+ (reconstituted) cell line. A 2-tailed unpaired t test was performed in I, but in this case, each KO was compared to the Huh7-ACE2 control cells. Used reagents are listed in detail in Table 1 . CoV, coronavirus; FKBP8, FK506 binding protein 8; HCoV, human ...