Journal: Nature Communications

Article Title: ZBP1 not RIPK1 mediates tumor necroptosis in breast cancer

doi: 10.1038/s41467-021-23004-3

Figure Lengend Snippet: a FVB/NJ mice at three weeks post-implantation with MVT-1 cells were treated weekly with vehicle or Necrostatin-1 (Nec-1; i.v .) until week 5. Left panel shows the representative images of H&E stained 5-week tumors of vehicle or Nec-1 treated mice. Scale bar, 2 mm. Right panel shows the percentage of tumor necrosis area (TN) of the total tumor area from mice at 5-week. Data are presented as mean values ± SEM. b Representative H&E and immunohistological images of phospho-MLKL (p-MLKL) antibody staining of tumor sections from mice implanted and treated as in Fig. a . Scale bar, 50 µm. c MVT-1 CRISPR/Cas9 control (CRISPR CT) or MVT-1 RIPK1 knock out (RIPK1 KO) cells were generated by using the CRISPR/Cas9 system. Left panel shows the representative H&E image of 4-week tumors from FVB/NJ mice implanted with the MVT-1 CRISPR CT or MVT-1 RIPK1 KO cells. Scale bar, 2 mm. Right panel shows the percentage of tumor necrosis area (TN) of the total tumor area after 4-weeks. Data are presented as mean values ± SEM. d Representative H&E and immunohistological images of p-MLKL antibody staining of 4-week tumor sections from mice implanted as in Fig. c . Scale bar, 50 µm. e Western blotting analysis of tumor lysates from mice implanted as in Fig. c using the indicated antibodies. Western blotting analysis representative of three independent experiments. Two-sided student’s t test was used to determine the statistical significance of differences between groups. Differences with P values < 0.05 were considered significant. Source data are provided as a Source Data file.

Article Snippet: MVT-1 inducible RIPK1 KO stable cells were generated using the Lenti-X tet-on 3G CRISPR-Cas9 system from Clontech (Takara Bio).

Techniques: Staining, CRISPR, Knock-Out, Generated, Western Blot

Journal: Nature Communications

Article Title: ZBP1 not RIPK1 mediates tumor necroptosis in breast cancer

doi: 10.1038/s41467-021-23004-3

Figure Lengend Snippet: a Western blotting analysis representative of three independent experiments. MVT-1 CRISPR/Cas9 control (CRISPR CT) or ZBP1 knock out (ZBP1 KO) tumor lysates were blotted using the indicated antibodies (upper panel). Tumor growth curve by measuring tumor volume of FVB/NJ mice implanted with MVT-1 CRISPR CT or ZBP1 KO cells (lower panel). b Left panel shows the representative images of H&E stained tumors at 5-weeks post-implantation as in Fig. a . Scale bar, 2 mm. Right panel shows the percentage of tumor necrosis area (TN) of the total tumor area from mice at 5-weeks post-implantation. Data are presented as mean values ± SEM. c Representative images of H&E and immunohistological stained sections with phospho-MLKL (p-MLKL) or cleaved caspase-3 (cl.Casp-3) antibodies of 5-week tumor sections from mice implanted as in Fig. a . Scale bar, 50 µm. d Western blotting analysis is representative of three independent experiments. 5-week tumor lysates from mice implanted as in Fig. a was determined by using the indicated antibodies. e Left panel shows the representative images of H&E stained lung sections from mice implanted as in Fig. a showing lung metastasis. Scale bar, 2 mm. Right panel shows the quantification of metastatic foci in lungs from mice at 5-week post-implantation. Data are presented as mean values ± SEM. Two-sided student’s t test was used to determine the statistical significance of differences between groups. Differences with P values < 0.05 were considered significant. Source data are provided as a Source Data file.

Article Snippet: MVT-1 inducible RIPK1 KO stable cells were generated using the Lenti-X tet-on 3G CRISPR-Cas9 system from Clontech (Takara Bio).

Techniques: Western Blot, CRISPR, Knock-Out, Staining