Extended Data Fig. 6: SARS-CoV-2 provokes senescence-like phenotype in hBOs.
a, RT-qPCR analysis of ACE2 gene expression in HDFs, HepG2, and human bronchial organoid (hBO). b, Schematic diagram of the genome (genomic-CoV2) and subgenomic (subgenomic-CoV2) RNAs and the positions of the RT-qPCR primers used (upper panel). hBOs were infected with SARS-CoV-2 at MOI of 0.1 for the indicated periods (lower panel). These cells were analyzed by RT-qPCR to determine the genomic and subgenomic SARS-CoV-2 RNAs (lower panel), gene expression of virus-induced cytokines and SASP factors (c), or by immunofluorescence analysis of KRT5 (basal cell marker) expression [red], p16INK4a expression [green], and DAPI [blue] (d), FoxJ1 (ciliated cell marker) expression [red], p16INK4a expression [green], and DAPI [blue] (e) or cleaved caspase-3 (C-casp 3) [red], SARS-CoV-2 spike protein (CoV2-S) [green], and DAPI [blue] (f) in Mock or SARS-CoV-2-infected cells (CoV2) on day 6 after SARS-CoV-2 infection. (Scale bars, 50 μm). The histograms indicate the percentages of p16INK4a expressing cells in KRT5 or FoxJ1 positive cell, and cleaved caspase-3 positive cell percentages in SARS-CoV-2 spike protein negative (CoV2-) or positive (CoV2 + ) cells. For all graphs, error bars indicate mean ± standard deviation (s.d.) of biological triplicate. Statistical significance was determined with two-tailed unpaired Student’s t test in (a), (d) and (e), and with two-way ANOVA followed by sidak’s multiple comparison test in (b), (c), and (f).
