Fig. 2: sgRNA screening of candidate TS genes revealed NF1, TSC1, and TβRII with metastasis-suppressive function. | Nature Communications

Fig. 2: sgRNA screening of candidate TS genes revealed NF1, TSC1, and TβRII with metastasis-suppressive function.

From: Loss of tumor suppressors promotes inflammatory tumor microenvironment and enhances LAG3+T cell mediated immune suppression

Fig. 2

A Schematic loss-of-function screening for TS genes in 4T1 mammary tumor metastasis (created with BioRender.com). B Lung nodule counts at 3 weeks (left) or 4 weeks (right) after tumor cell injection and C Lung nodule counts with different sizes from immune-deficient mice received tumor cell injection. The number and size of tumor nodules on lung surfaces were counted under a dissecting microscope. n = 8 per group. D Representation of sgRNA library at different stages of tumor growth and metastasis. Number of sgRNA species in cells before transplantation, different stages of primary tumor, lymph node, and lung during tumor evolution. E Pie charts for the most abundant sgRNAs in the lung after 2, 3, and 4-week sgRNA library-mediated cell injection. F Enriched sgRNAs in the lung nodules at 3 and 4 weeks after cell transplantation. Individual tumor nodules were taken out from the lungs, PCR amplified the sgRNA sequence, and examined through Sanger sequencing. G Dynamic evolution of sgRNAs indicated during tumor growth and metastasis. All sgRNAs with ≥2% of total reads are plotted individually. The remaining lung indicates most of the nodules were picked out before the evaluation. n = 8 per group. All bar graphs show mean ± s.d. Statistical significance was determined by two-tailed Student’s t-test for (C).

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