Fig. 4

LINC00301 facilitates regulatory T cells infiltration in tumors isolated from mouse NSCLC cell lines planted A/He or DBA/2 mice, respectively. a, b Representing images for CyTOF analysis of tumors by PhenoGraph analysis, identifying 22 clusters, colored by cluster identification numbers, and plotted by tSNE1 and tSNE2, clarified various clusters with obvious alterations between groups. c Flow cytometry gating method used in the analysis of CD3⁺/CD4⁺/CD25⁺ regulatory T cells infiltration was presented. Shown is a pLenti-CMV-vector tumor cell preparation. Following a preliminary assessment of a lymphocyte gate according to forward versus side scatter, feasible cells (FITC positive) were identified as CD3⁺ cells. The resulting CD3⁺ population was following gated according to positivity for CD4 ⁺/CD25⁺. The frequencies of cells in these populations were normalized by background values and then submitted and evaluated directly. d Flow cytometry measurement of percentage of CD3⁺/CD4⁺/CD25⁺ Treg cells. e Representative images of CD4 and CD25 of tumors isolated from sh-NC, sh-1, pLenti-CMV-vector, or pLenti-CMV-LINC00301 treated LA-4 and KLN 205 cells planting in A/He or DBA/2 mice, respectively (n = 5 animals for each group). f Representative images of CD8 of tumors isolated from sh-NC, sh-1, pLenti-CMV-vector, or pLenti-CMV-LINC00301-treated LA-4 and KLN 205 cells planting in A/He or DBA/2 mice, respectively (n = 5 animals for each group). Scale bar = 50 μm, p values were established by unpaired two-tailed Student’s t-test