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. 2022 Nov 4;141(7):766–786. doi: 10.1182/blood.2022017046

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Figure 5. — C1Q⁺Molm13 cells are highly infiltrative toward tissue fibroblasts. (A) Flow cytometry sorting of Molm13 based on expression of C1Q. (B-C) Transwell migration assay of Molm13^C1Q+ and Molm13^C1Q−, or Molm13 cocultured with HaCaT, skin fibroblasts, or GI fibroblasts. (D) qPCR analysis of C1QA in Molm13 cells upon depletion of C1QA through shRNA (shC1QA-1 and shC1QA-2). (E) Transwell migration assay of Molm13 cells upon C1QA depletion. (F) NSG mice were injected with one million of Molm13^C1Q+ or Molm13^C1Q− cells and metastasis nodules in skin and intestine are numbered and shown. (G) Number or size of skin or GI nodules were calculated. (H-I) qPCR analysis of C1QA (H) and TGF-β1, TGF-β2, and TGF-β3 (I) in Molm13^C1Q+ and Molm13^C1Q− cells. (J) qPCR analysis of TGF-β1, TGF-β2, and TGF-β3 in Molm13^C1Q+ cells cocultured with or without fibroblasts. (K) Western blot analysis of TGF-β1 of Molm13^C1Q+ and Molm13^C1Q− cells cocultured with or without fibroblasts. (L) Transwell migration assay of Molm13^C1Q+ cells upon skin fibroblast coculture and TGF-β1 inhibitor galunisertib treatment.

Figure 5. — C1Q⁺Molm13 cells are highly infiltrative toward tissue fibroblasts. (A) Flow cytometry sorting of Molm13 based on expression of C1Q. (B-C) Transwell migration assay of Molm13^C1Q+ and Molm13^C1Q−, or Molm13 cocultured with HaCaT, skin fibroblasts, or GI fibroblasts. (D) qPCR analysis of C1QA in Molm13 cells upon depletion of C1QA through shRNA (shC1QA-1 and shC1QA-2). (E) Transwell migration assay of Molm13 cells upon C1QA depletion. (F) NSG mice were injected with one million of Molm13^C1Q+ or Molm13^C1Q− cells and metastasis nodules in skin and intestine are numbered and shown. (G) Number or size of skin or GI nodules were calculated. (H-I) qPCR analysis of C1QA (H) and TGF-β1, TGF-β2, and TGF-β3 (I) in Molm13^C1Q+ and Molm13^C1Q− cells. (J) qPCR analysis of TGF-β1, TGF-β2, and TGF-β3 in Molm13^C1Q+ cells cocultured with or without fibroblasts. (K) Western blot analysis of TGF-β1 of Molm13^C1Q+ and Molm13^C1Q− cells cocultured with or without fibroblasts. (L) Transwell migration assay of Molm13^C1Q+ cells upon skin fibroblast coculture and TGF-β1 inhibitor galunisertib treatment.