Fig. 2

LINC01977 exerts oncogenic roles in vitro and in vivo. A Number of proliferating LINC01977-silenced A549 cells, as determined by the EdU incorporation assay. Cells transfected with scramble were used as control. DAPI was used to stain the nuclei. Scale bars: 200 μm. B Different cell cycle phases of LINC01977-silenced A549 cells. Cells were stained using propidium iodide (PI), and the percentages were determined by flow cytometry. C The apoptosis of LINC01977-silenced A549 cells. Cells were stained using PI and annexin V, and the percentages were determined by flow cytometry. D The invasion, migration of LINC01977-silenced A549 cells assessed by wound healing and transwell assays (up panel). The proliferation ability of LINC01977-silenced A549 cells assessed by spheroid formation and colony formation assay (down panel). E Schematic representation of xenograft nude mouse model. F Representative images of mice-bearing tumors derived from xenograft nude mouse model. G qRT-PCR showing mRNA expression levels of LINC01977 in tumor tissues obtained from subcutaneous model. H H&E stained and ZEB1 IHC of serial sections from mice-bearing tumors. H&E, scale bars: 100 μm; IHC, scale bars: 20 μm. Tumor weight (I) and tumor volume (J) were assessed. Data point in represent (mean ± SEM) are shown from n = 6 biologically independent samples by two-sided unpaired t test. K Schematic representation of xenograft metastasis model. L Representative images of IVIS detection and metastatic loci in the lung derived from mouse model. M H&E stained and ZEB1 IHC detection of serial sections from lung metastasis nude mouse model. H&E, scale bars: 100 μm; IHC, scale bars: 20 μm. Data in A–D are representative of three independent experiments and presented as mean ± S.D., A–D (n = 3), G, I and J (n = 6) biologically independent samples, and the P value was determined by a two-tailed unpaired Student’s t test and one-way ANOVA, respectively