Figure 2: miR-1246 and miR-1290 contribute towards transformation and lung tumorigenesis. | Nature Communications

Figure 2: miR-1246 and miR-1290 contribute towards transformation and lung tumorigenesis.

From: Tumour-initiating cell-specific miR-1246 and miR-1290 expression converge to promote non-small cell lung cancer progression

Figure 2

(a) Sphere-formation assay for tumoursphere cells treated with either miR-1246 knockdown (zip1246) or miR-1290 knockdown (zip1290). A total of 500 cells were seeded into each 10-cm dish. Spheres containing >50 cells were counted on day 13. Scale bar, 200 μm. (b) Limiting dilution analysis of sphere-formation efficiency for tumoursphere cells treated with either zip1246 or zip1290. 50, 150 and 500 cells were plated; n=3. (c,d) Images (c) and quantitative analysis of volume and mass (d) of tumours formed 28 days after subcutaneous transplantation of 100,000 tumoursphere cells that were treated with either zip1246 or zip1290; n=6. (e) Limiting dilution analysis of tumour initiation by tumoursphere cells treated with either zip1246 or zip1290. In all, 100 and 2,000 cells were transplanted subcutaneously and tumour formation was evaluated 90 days later. The representative mouse images on transplantation (left) and the number of xenografted tumours/number of injections are shown; n=8. (f) qRT–PCR analysis of miR-1246 and miR-1290 expressions in HEK293 infected with either pre-miR-1246 (pre1246) or pre-miR-1290 (pre1290) overexpression. (g,h) Colony-formation assay (g) and quantification (h) in adherent conditions of HEK293 treated with either pre1246 or pre1290 overexpression. In all, 100 cells were plated; n=3. (i) Xenograft tumour-formation efficiency of HEK293 treated with either pre1246 or pre1290. A total of 100,000 cells were transplanted subcutaneously and tumour formation was evaluated 60 days later; n=6. (j,k) Soft-agar colony-formation assay (j) and quantification (k) in NuLi-1 treated with either pre1246 or pre1290. A total of 10,000 cells were plated and the colonies were stained by INT after 28 days. (l) Xenograft tumour-formation efficiency of NuLi-1 treated with either pre1246 or pre1290. A total of 1,000,000 cells were transplanted subcutaneously and tumour formation was evaluated 60 days later; n=6. (m) Xenograft tumour formation of CD166− tumour cells from xenograft tumour treated with pre1246 or pre1290. A total of 100,000 and 250,000 cells were transplanted subcutaneously and tumour formation was evaluated 90 days later; n=10. All error bars represent±s.e.m. and statistical significance was calculated using Student’s t-test; *P<0.05, **P<0.01.

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