Fig. 4: NLRP7 knockout induces apoptosis of embryonic cells in blastoids.

A Representative immunofluorescent images of wild type and NLRP7 knockout blastoids at day 3 and day 4 for the DNA damage marker γH2A.X (red) and the epiblast marker OCT4 (green). The nuclei were counterstained with DAPI and are shown in blue. Elevated DNA damage is detected in the NLRP7^−/− blastoids, and most of the γH2A.X⁺ cells co-express OCT4. Scale bar, 100 μm. B Quantification of the γH2A.X-positive cells in the immunofluorescent staining (WT Blastoids, n = 13 biological replicates; NLRP7^−/− blastoids, n = 11 biological replicates) in (A). Box plots show median (center line), 25th and 75th percentiles (bottom and top of the box, respectively), and minimum and maximum values (bottom and top whisker, respectively). Data are shown as mean ± SEM. Student’s t-test. C Representative immunofluorescent images of wild type and NLRP7 knockout blastoids at day 4 for the Apoptosis marker active caspase 3 (red) and the epiblast marker OCT4 (green). The nuclei were counterstained with DAPI and are shown in blue. Elevated apoptosis is detected in the NLRP7^−/− blastoids, and most of the active caspase 3⁺ cells co-express OCT4. Scale bar, 100 μm. D Quantification of the active caspase 3⁺ cells in the immunofluorescent staining (WT Blastoids, n = 11 biological replicates; NLRP7^−/− blastoids, n = 10 biological replicates) in (C). Box plots show median (center line), 25th and 75th percentiles (bottom and top of the box, respectively), and minimum and maximum values (bottom and top whisker, respectively). Data are shown as mean ± SEM. Student’s t-test.