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HEP_25895_sm_SuppFig1.tif1059KSupporting Information Figure 1. Full-length of human lncRNA MVIH gene cloning. (A) Left: Agarose gel electrophoresis of nested PCR products from the 5′-RACE procedure and the 3′-RACE procedure. Molecular weight markers (base pairs) are indicated on the right. The major PCR product is marked by an arrow on the left. Right: Sequencing of second-round PCR products revealed the boundary between the universal anchor primer and lncRNA MVIH sequences. The guanine marked by a vertical line indicates a putative transcriptional start site or a putative transcriptional end site. The arrows indicate the transcriptional directions. (C) Nucleotide sequence of the full-length human lncRNA MVIH gene. Sequence added based on reference sequence in NCBI database is indicated in red.
HEP_25895_sm_SuppFig2.tif8136KSupporting Information Figure 2. Codon substitution frequency scores (CSFs) of lncRNA-MVIH and RPS24. The whole transcription region is divided to some blocks by chromosome positions of exon, intron of RPS24 and MVIH. The average score of each block was taken as the CSFs of corresponding block. The blue bars represent exons of RPS24 and the green bar represents the transcript of lncRNA MVIH. The black lines represent regions un-transcribed. TSS, transcription start site. TES, transcription end site.
HEP_25895_sm_SuppFig3.tif12783KSupporting Information Figure 3. (A) Expression pattern of lncRNA MVIH in HCC cell lines and normal cell line (LO2). (B) The expression of the lncRNA MVIH in Huh7 and HCCLM3 cells transfected with a lentivirus encoding the lncRNA MVIH (Huh7-lncRNA-MVIH and HCCLM3-lncRNA-MVIH) or a puromycin control (Negative control, Huh7-NC and HCCLM3-NC) was quantified by RT-PCR. The data are shown as the means ± standard deviation of three independent biological replicates. The statistical difference was analyzed by Two-sample t-test. (C) Representative photographs of luciferase signals in 1 week and 5 weeks were taken by an IVIS system after injection with HCCLM3 cells through the splenic hilum.
HEP_25895_sm_SuppFig4.tif2993KSupporting Information Figure 4. (A) RIP experiments were performed using an antibody against EZH2 on extracts from SMMC-7721 cells and one fresh HCC tissue. The purified RNA was used for q RT-PCR using the SYBY-green method, and the enrichment of the lncRNA MVIH was normalized to the input. The lncRNA HEIH, which was previously revealed to associate with EZH2, was applied for a positive control. The data are shown as the means ± S.D. The statistical difference was analyzed by Two-sample t-test. (B) Expression of key genes in glycolysis was detected using q RT-PCR. Huh7 and HCCLM3 transfected with Lv-lncRNA-MVIH showed no significant alteration of glycoltic markers expression compared with control cells. (C) The production of lactate acid in Huh7 and HCCLM3 cells transfecting LV-lncRNA-MVIH or LV-NC was measured by lactate acid detection kit (JianCheng, NanJing, China) according to manufacturer's instructions.The data were showed as mean ± S.D. The statistical difference was analyzed by Two-sample t-test.
HEP_25895_sm_SuppInfo.doc150KSupporting Information

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