Fig. 6: Human LINC01271 is the human ortholog of MaTAR25.

a All potential human orthologs of MaTAR25 (hMaTAR25) were identified based on conservation of genomic location (synteny). RNA-seq data from The Cancer Genome Atlas (TCGA) was analyzed to evaluate the expression status of all potential hMaTAR25 candidates by comparison of 1128 TCGA breast tumor datasets to 113 normal breast tissue controls. Fold change and statistical significance were calculated using the Wilcoxon rank test; P values were adjusted using the Benjamini & Hochberg method by DESeq2³⁰. b Attempted rescue of 4T1 MaTAR25 KO cells upon ectopic expression of two transcript isoforms of LINC01270 (LINC01270.1, and LINC01270.2), or LINC01271 in cell viability assays. The mean cell numbers of three independent replicates of 4T1 control1, MaTAR25 KO1, MaTAR25 KO1 with GFP, LINC01270.1 clone1-5, LINC01270.2 clone1-5, and LINC01271 clone1-3 are shown ± SD (n = 3) *P < 0.05 (paired Student’s t test; two-tailed). c 4T1 MaTAR25 KO cells with ectopic expression of GFP was used as a control to assess rescue in a cell invasion assay. The mean relative cell invasion of two independent replicates of 4T1 control, MaTAR25 KO1, MaTAR25 KO1 with GFP clone1-2, LINC01271 ectopic expression clone1-2 is shown ± SD (n = 2 independent experiments). Ectopic expression of LINC01271 can rescue the MaTAR25 KO cell invasion phenotype. d RNA expression level of Tns1 was determined in MaTAR25 KO1 cells ectopically expressing LINC01270.1, LINC01270.2, or LINC01271 by qRT-PCR. Data are presented as mean values ± SD (n = 3 independent experiments). *P < 0.05 (paired Student’s t test; two-tailed). The protein level of mTns1 was also examined in MaTAR25 KO cells with ectopic expression of LINC01271 by immunoblot analysis. e Three different ASOs targeting LINC01271 were used to independently knockdown LINC01271 in MDA-MB-231 LM2 cells. Left panel: the knockdown efficiency is calculated and data are shown as mean values ± SD (n = 2 independent experiments) by qRT-PCR after 24 h treatment of ASOs. Right panel: the mean cell numbers of three independent cell counting experiments of MDA-MB-231 LM2 mock-treated control cells, cells treated with scrambled ASO, and cells treated with three different LINC01271 ASOs are shown ± SD (n = 3). *P < 0.05 (paired Student’s t test; two-tailed).