Fig. 6

RGS6 knockout reduced the stemness and self-renewal capacity of AEC2s. A Primary AEC2s morphology was observed under light microscope. B Immunofluorescence staining showed the purity of AEC2s. Cell nucleus was stained by DAPI (blue fluorescence). AEC2s were stained by anti-SFTPC antibody (green fluorescence). C Organoids came to being at day 3, characterized as two or three cells grouped to form a ring structure (as shown by the black arrow). Alveolar organoids formation from RGS6^−/− mice was slower compared to that from WT mice. D Alveolar organoids observation at day 7. E Alveolar organoids could still form and develop after passaging in WT group, which was rarely seen in RGS6^−/− group. F The internal structure of alveolar organoids was shown under high magnification microscope. G Immunofluorescence staining showed AEC2s could differentiate into AEC1s in alveolar organoids. Cell nucleus was stained by DAPI (blue fluorescence). AEC2s were stained by anti-SFTPC antibody (green fluorescence). AEC1s were stained by anti-PDPN antibody (red fluorescence). H Alveolar organoids counting at day 9 in different groups. Scale bars were shown in the pictures. WT, the primary AEC2s isolated from lung tissue of wild type mice and cultured in Matrigel. RGS6^−/−, the primary AEC2s isolated from lung tissue of RGS6^−/− mice and cultured in Matrigel. Data are shown as mean ± SEM. ^***P < 0.001 vs WT group