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. 2021 Nov 16;12:6610. doi: 10.1038/s41467-021-26762-2

Fig. 4. Reverse polarity organoids for efficient SARS-CoV-2 infection.

Fig. 4

a Schematic showing the generation of reverse polarity gastric organoids (RP-GOs). b Gastric organoids with normal (inner) polarity and large lumen. Immunofluorescence panel showing f-actin (F-ACT) in red, zonula occludens-1 (ZO-1) in violet, β-4 integrin (β-4 INT) in red, and nuclei in blue (Hoechst). Scale bar 50 μm. c RP-GOs showing an almost absent lumen. Immunofluorescence panel showing f-actin (F-ACT) in red, zonula occludens-1 (ZO-1) in violet, β-4 integrin (β-4 INT) in red, mucin 5AC (MUC5AC) in green and nuclei in blue (Hoechst). Scale bar 50 μm. d, e SARS-CoV-2 receptors, angiotensin-converting enzyme 2 (ACE2) and transmembrane protease serine 2 (TMPRSS2) absolute RNA-seq expression in gastric tissues (left) and gastric organoids (right). Mean ± SD (n = 3 for tissues biological replicates, n = 3 for organoids). CPM count per million. Horizontal lines above the bar plots highlight DEGs between the indicated conditions. f Immunofluorescence panel showing ACE2 in red, TMPRSS2 in green and nuclei in blue (Hoechst) in fetal and pediatric stomach biopsies. Scale Bar 50 µm (main figures) and 30 µm (enlargement). g Immunofluorescence panel showing ACE2 in red, TMPRSS2 in green, and nuclei in blue (Hoechst) in fetal, pediatric, and adult gastric organoids with reverse polarity. Scale bar 30 μm (main figures) and 10 µm (enlargement).