Fig. 1: NeoCoV and PDF-2180 use ACE2 but not DPP4 for efficient entry.
From: Close relatives of MERS-CoV in bats use ACE2 as their functional receptors
a,b, Phylogenetic analysis of merbecoviruses (grey) based on whole-genome nucleotide sequences (a) and S1 amino acid sequences (b). NL63 and 229E were set as outgroups. Host and receptor use are indicated. For the scale bars, 0.5 represents two nucleotide substitutions per site. c, Simplot analysis showing the whole-genome nucleotide sequence similarity of three merbecoviruses compared with MERS-CoV. The boundaries of regions encoding MERS-CoV proteins are indicated at the top. The box delineated by a dashed line underscores the divergence of the S1 subunit. T/t, transition/transversion ratio. d, Pseudotyped S virus entry efficiency of six merbecoviruses in BHK-21 cells transiently expressing hACE2, hDPP4 or hAPN. The dashed lines indicate the baseline of background signals (mean values of vector-only groups). Data are mean ± s.e.m., representative of three independent experiments. n = 3 biologically replicates. Statistical analysis was performed using two-tailed unpaired Student’s t-tests. PSV, pseudotyped virus. e, The entry efficiency of NeoCoV S pseudotyped virus in HEK293T stable cell lines expressing different bat ACE2 orthologues as indicated by luciferase activity. Data are mean ± s.d. of biological triplicates examined over three independent infection assays. f,g, NeoCoV, PDF-2180 and MERS-CoV S-mediated cell–cell fusion analysis based on DSP assays in HEK293T cells stably expressing the indicated receptors. TPCK-trypsin, TPCK-treated trypsin treatment (10 μg ml−1). eGFP intensity (f) and live-cell Renilla luciferase activity (g) are shown. Data are mean ± s.d. of n = 4 biologically independent cells. Three independent cell–cell fusion assays are shown. Statistical analysis was performed using two-tailed unpaired Student’s t-tests. For f, scale bars, 200 μm. h, Pseudotyped virus entry efficiency of six merbecoviruses in HEK293T cells stably expressing the indicated bat ACE2 or DPP4 orthologues. Data are representative results of three independent experiments and plotted by the mean of the biological triplicate. RLU, relative light units. **P < 0.01, ****P < 0.001.
