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. 2019 Nov 21;9:17319. doi: 10.1038/s41598-019-53944-2

Figure 1.

Figure 1

EVA1A-AS is expressed in HepG2 cells, but not in normal hepatocytes and suppresses EVA1A expression. HepG2 cells were infected with lentivirus carrying shTHOC5 (shTHOC5) or sh control (shCr) for three days. (A) Cell lysates were applied for THOC5 specific immunoblot. GAPDH specific immunoblot was performed as loading control. RNAs were isolated from a sister culture and applied for THOC5, SPC25, ID1 specific qRT-PCR. (B) RNA sequence data of THOC5. SeqMonk was used to quantitate and visualize the data. Read density is shown as reads per million (RPM). (C) Predicted EVA1A-AS (NONHSAG028257.2) (+strand) and EVA1A (−strand) in HepG2 cells infected with lentivirus carrying shRNA control or shTHOC5. Total RNA-seq datasets from human liver tissue (ENCFF098AZJ) were generated by the ENCODE Consortium and were aligned to the reference human genome (GRCh38). Black box: exon. (D) RNAs were isolated from HepG2 treated with siCr, siEVA1A-AS-1, or siEVA1A-AS-2 and were supplied for EVA1A, EVA1A-AS or GAPDH specific semi-quantitative RT-PCR. []: number of PCR cycles. (E) RNAs were isolated from HepG2 cells treated with siCr, or siEVA1A and supplied for EVA1A-AS, EVA1A or GAPDH specific semi-quantitative RT-PCR. []: number of PCR cycles. (F) RNAs were isolated from sister culture of (D) and applied for EVA1A-AS, EVA1A specific q-RT-PCR. Three independent experiments were performed. Bars represent +/−SD. (G) Cell extracts from a sister culture of (D) were applied for EVA1A and GAPDH specific immunoblot.