Journal: Aging

Article Title: LncRNA SH3BP5-AS1 promotes hepatocellular carcinoma progression by sponging miR-6838-5p and activation of PTPN4.

doi: 10.18632/aging.205811

Figure Lengend Snippet: Figure 1. SH3BP5-AS1 and miR-6838-5p have a relationship in HCC. (A, B) HCC cells and tissues showed increased SH3BP5-AS1 expression. (C) The patients’ advanced tumor stage was indicated by the elevated SH3BP5-AS1. (D) The position of SH3BP5-AS1 was identified using subcellular fractionation analysis. (E) The binding sites of SH3BP5-AS1 and miR-6838-5p. (F) MiR-6838-5p inhibited the activity of the luciferase gene in SH3BP5-AS1 wild-type (SH3BP5-AS1-WT), but not in SH3BP5-AS1 mutant (SH3BP5-AS1-MUT). (G) SH3BP5- AS1 prevented MiR-6838-5p from being expressed. (H) MiR-6838-5p mimics decreased SH3BP5-AS1 expression. (I) In HCC tissues, miR-6838- 5p expression was markedly downregulated. (J) A negative correlation was observed between SH3BP5-AS1 and miR-6838-5p expression.

Article Snippet: Cell lines and cell culture Human HCC cell lines (SMCC7721, Huh7, HepG2, and Hep3B) and normal hepatic epithelial cell LO2 cell lines were obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Expressing, Fractionation, Binding Assay, Activity Assay, Luciferase, Mutagenesis

Journal: Aging

Article Title: LncRNA SH3BP5-AS1 promotes hepatocellular carcinoma progression by sponging miR-6838-5p and activation of PTPN4.

doi: 10.18632/aging.205811

Figure Lengend Snippet: Figure 2. The impact of SH3BP5-AS1 was diminished by miR-6838-5p. (A) SH3BP5-AS1 expression in HCC cells transfected with pcDNA-SH3BP5-AS1 and miR-6838-5p (or mim-ctrl) was determined by qRT-PCR assay. (B, C) CCK-8 and colony formation assays were used to investigate the effect of the SH3BP5-AS1/miR-6838-5p axis on the proliferation capacity of the HCC cells. (D–F) Wound healing and transwell assays were used to assess the impact of the SH3BP5-AS1/miR-6838-5p axis on the migration and invasion capacity of the HCC cells.

Article Snippet: Cell lines and cell culture Human HCC cell lines (SMCC7721, Huh7, HepG2, and Hep3B) and normal hepatic epithelial cell LO2 cell lines were obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Expressing, Transfection, Quantitative RT-PCR, CCK-8 Assay, Migration

Journal: Aging

Article Title: LncRNA SH3BP5-AS1 promotes hepatocellular carcinoma progression by sponging miR-6838-5p and activation of PTPN4.

doi: 10.18632/aging.205811

Figure Lengend Snippet: Figure 3. PTPN4 was a functional target of miR-6838-5p. (A) The PTPN4 mRNA 3-UTR’s miR-6838-5p binding location. (B, C) MiR-6838- 5p prevented PTPN4’s mRNA and protein from being expressed. (D) Luciferase activity was significantly decreased when PTPN4-WT and miR- 6838-5p were co-transfected. (E) The qRT-PCR technique was used to measure the mRNA level of PTPN4 in the HCC cells. (F, G) PTPN4 silencing prevented the miR-6838-5p inhibitor’s promotion effect on HCC cells’ growth. (H, I) The cell scratch and transwell assays were used to identify the migration and invasion capacity of HCC cells.

Article Snippet: Cell lines and cell culture Human HCC cell lines (SMCC7721, Huh7, HepG2, and Hep3B) and normal hepatic epithelial cell LO2 cell lines were obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Functional Assay, Binding Assay, Luciferase, Activity Assay, Transfection, Quantitative RT-PCR, Migration

Journal: Aging

Article Title: LncRNA SH3BP5-AS1 promotes hepatocellular carcinoma progression by sponging miR-6838-5p and activation of PTPN4.

doi: 10.18632/aging.205811

Figure Lengend Snippet: Figure 5. SH3BP5-AS1 accelerated HCC cell proliferation in vivo. (A–C) Tumor xenograft assay in the indicated HCC cells.

Article Snippet: Cell lines and cell culture Human HCC cell lines (SMCC7721, Huh7, HepG2, and Hep3B) and normal hepatic epithelial cell LO2 cell lines were obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: In Vivo, Xenograft Assay

Journal: Aging

Article Title: LncRNA SH3BP5-AS1 promotes hepatocellular carcinoma progression by sponging miR-6838-5p and activation of PTPN4.

doi: 10.18632/aging.205811

Figure Lengend Snippet: Figure 6. HIF-1α regulated SH3BP5-AS1 transcription by directly binding to its promoter. (A) The mRNA level of SH3BP5-AS1 in the hypoxic HCC cells was measured using a qRT-PCR experiment. (B) HIF-1α binding motif. (C) Possible HIF-1α binding location in the SH3BP5- AS1 promoter. (D) CHIP-qPCR assay. (E) Dual luciferase reporter assay in the HCC cells. (F) qRT-PCR experiment in the HCC cells.

Article Snippet: Cell lines and cell culture Human HCC cell lines (SMCC7721, Huh7, HepG2, and Hep3B) and normal hepatic epithelial cell LO2 cell lines were obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Binding Assay, Quantitative RT-PCR, ChIP-qPCR, Luciferase, Reporter Assay

Journal: Cell Death & Disease

Article Title: Non-canonical role of wild-type SEC23B in the cellular stress response pathway

doi: 10.1038/s41419-021-03589-9

Figure Lengend Snippet: A Non-malignant thyroid Nthy-ori 3-1 cells were stably transduced with EGFP, wild-type SEC23B , and mutant p.Val594Gly SEC23B (the latter two fused with EGFP). Nthy- SEC23B -WT and Nthy- SEC23B- V594G cells show nuclear expression of SEC23B. Nthy-EGFP cell line is used as a control to show non-specific GFP subcellular localization. Blue, DAPI; green, SEC23B-EGFP or EGFP. Scale bars, 25 μm. B Protein levels after nuclear–cytoplasmic fractionation in stably transduced Nthy-ori 3-1 cells detects endogenous and EGFP-tagged SEC23B in the nuclear compartment. Nthy-EGFP cell line is used as a control to indicate baseline endogenous levels of SEC23B. PARP1, nuclear marker; GAPDH, cytoplasmic marker; CANX, endoplasmic reticulum marker. C Protein levels after nuclear–cytoplasmic fractionation in parental HEK293 cells detects endogenous SEC23B in the nuclear compartment. PARP1 and Lamin A/C, nuclear markers; GAPDH, cytoplasmic marker; CANX, endoplasmic reticulum marker.

Article Snippet: HEK293 cells (originally purchased from the American Type Culture Collection in 2011 and obtained in 2014 from the Cleveland Clinic Lerner Research Institute Cell Culture Core) were cultured in Dulbecco’s modified Eagle’s medium supplemented with 10% FBS and 1% penicillin and streptomycin.

Techniques: Stable Transfection, Transduction, Mutagenesis, Expressing, Control, Fractionation, Marker

Journal: Cell Death & Disease

Article Title: Non-canonical role of wild-type SEC23B in the cellular stress response pathway

doi: 10.1038/s41419-021-03589-9

Figure Lengend Snippet: A Truncation mutant constructs generated to characterize the bioinformatically top scoring NLS sequences and the single predicted NES sequence. The full-length SEC23B protein (Uniprot Q15437, SC23B_HUMAN) is depicted in the top panel. aa amino acids. B Confocal microscopic images of HEK293 cells transiently transfected with each of the constructs. The corresponding localization of the SEC23B protein is summarized in A . The predicted NES overlaps with the predicted NLS1 sequence. Scale bars, 8 μm.

Article Snippet: HEK293 cells (originally purchased from the American Type Culture Collection in 2011 and obtained in 2014 from the Cleveland Clinic Lerner Research Institute Cell Culture Core) were cultured in Dulbecco’s modified Eagle’s medium supplemented with 10% FBS and 1% penicillin and streptomycin.

Techniques: Mutagenesis, Construct, Generated, Sequencing, Transfection