a Illustration of the location of RNA capture probes (30?k) and lncRNA transcribed from your antisense strand of the sequence around exon 1 of the gene (chr9: 21,989,178-21,994,898 in the human genome GRCh37/hg19). (c) A fragment deletion in exon 1 was detected by PCR (top chart) in HEK293T or HCT116 genes in HCT116 and HEK293T cells whose ARE-containing elements in exon-1 were homogenously deleted in the qRT-PCR analysis (bottom chart). Pooled ARE-KO-negative subclones were used as a wild-type (WT) control. (d) Detection of the P16, P15, and P14 proteins in HEK293T cells in Western blot analyses. 12943_2020_1150_MOESM4_ESM.docx (653K) GUID:?86A68C7D-72DE-4011-8EA6-B29F6AAA0F19 Additional file 5 Figure S3. expression decreased mRNA-AUF1 binding. (a) AUF1 directly bound to and mRNA in HCT116 cells in the AUF1-RIP-PCR. (b) overexpression decreased and mRNA-AUF1 conversation in by the AUF1-RIP-qPCR. (c) An illustration of how the competitive AUF1-binding protects and mRNA from your decay. AUF1 complexes were drawn as dimers based on the reports that AUF1 isoforms (p37, p40, p42, and p45) could form functional dimers [25, 26]. 12943_2020_1150_MOESM5_ESM.docx (341K) GUID:?547288CE-36D4-43C2-818F-8C8013459469 Additional file 6 Figure S4. Genome-wide analyses of transcriptome by RNA sequencing for HCT116 cells with and without overexpression and/or AUF1 downregulation. The HCT116 cells with stable overexpression were transfected with siRNAs (siAUF1) for 72?h, and then harvested for RNA sequencing. The number of genes with ?2 fold changes (UP, upregulated; Down, downregulated) for different types of RNAs were labeled. Western blot analysis for the determination of downregulation by siRNAs was inserted into the top chart. Two samples were sequenced for each group. 12943_2020_1150_MOESM6_ESM.docx (234K) GUID:?0FCDF77A-F764-47D2-BE21-BF7CF75AF959 Additional file 7 Table S2. Function annotations for and using the publicly available transcriptome databases for human malignancy cell lines in the CCLE project. (A) All 1037 cell lines; N8-Acetylspermidine dihydrochloride (B) 224 cell lines without the allele deletion (relative copy number? ?0). 12943_2020_1150_MOESM9_ESM.docx (333K) GUID:?981CD035-58E5-4623-98AD-FC7A59D05060 Additional file 10 Figure S6. Comparison of the levels of expression in colon tissues from malignancy and noncancer patients. (a) The expression status of in colon cancer (CC), paired surgical margin (SM), and normal colon biopsy (Normal) tissues from noncancer patients by qRT-PCR. (b) The level of expression in and mRNA (by qRT-PCR) in expression level (by qRT-PCR) in colon cancer (CC) and surgical margin (SM) tissue samples from patients with different clinicopathological characteristics 12943_2020_1150_MOESM11_ESM.docx (16K) GUID:?271CC986-08A5-48A6-ABBE-CA71D395DF81 Additional file 12 Figure S7. Comparisons of the levels of mRNA (by qRT-PCR) with those of and lncRNA in colon cancer tissues (CCs). 12943_2020_1150_MOESM12_ESM.docx (150K) GUID:?A3030D6F-AFF4-4669-A1E7-C1AEE3AD3D19 Additional file 13 Table S5. Comparison of the expression level (by RT-PCR) in colon cancer (CC) and surgical margin (SM) tissue samples from patients with different clinicopathological characteristics 12943_2020_1150_MOESM13_ESM.docx (17K) GUID:?9D74391F-00D3-4AA4-9C68-404F9998232B Additional file 14 Table S6. Comparison of the mRNA level (by qRT-PCR) in colon cancer (CC) and surgical margin (SM) tissue samples from patients with different clinicopathological characteristics 12943_2020_1150_MOESM14_ESM.docx (17K) GUID:?459EABB7-540E-428A-AF75-78152348B95E Additional file 15 Table S7. Comparison of and coexpression in colon cancer (CC) and surgical margin (SM) tissue samples from patients with different clinicopathological characteristics 12943_2020_1150_MOESM15_ESM.docx (18K) GUID:?89F5E0CE-4203-4FAC-B062-BE132B99847F Additional file 16 Physique S8. Graph of the gene in the locus. (A) CpG islands within the gene. (B) The N8-Acetylspermidine dihydrochloride transcription and active histone modification status in the chromatin upstream of the gene in 7 cell lines from ENCODE. (C) Transcription factors binding to numerous fragments round the gene from ENCODE. The RNA polymerase II (POLR2A) N8-Acetylspermidine dihydrochloride is usually highlighted in reddish lines. (D) The conservation status of various ITGAL fragments among vertebrates (adapted from your UCSC website). 12943_2020_1150_MOESM16_ESM.docx (291K) GUID:?C9DD37BE-071E-4A7D-B5FC-9F0F6FE1CEE5 Additional file 17 Figure S9. Repression of expression in gastric malignancy cells by designed locus contains crucial tumor suppressors and a lncRNA gene gene were screened by with its promoter in the antisense strand of the fragment near exon 1b in human cells. The mature is usually a three-exon linear cytoplasmic lncRNA (1043-nt), including an AU-rich element (ARE) in exon 1. decreases AUF1-RNA interaction and then increases expression by competitively binding to AUF1 P37 and P40 isoforms. Interestingly, significantly promoted the proliferation of malignancy cells and tumor formation in NOD-SCID mice in a and lncRNAs were significantly upregulated compared with the paired normal tissues. Conclusion A novel lncRNA, gene, promotes colon cancer development upregulating the expression of oncogenic and (locus at chromosome 9p21 [1]. P16 and P15 proteins target CDK4/6 through.
