Journal: bioRxiv

Article Title: The RNA-binding protein landscapes differ between mammalian organs and cultured cells

doi: 10.1101/2022.02.10.479897

Figure Lengend Snippet: (A) Venn diagram showing the number of shared and organ-specific RBPs identified. (B) Normalized signal sum of identified RBPs in ex vivo eRIC eluates (upper panel) and the corresponding input samples (middle panel) for each organ analyzed. Center lines indicate the median, box borders represent the interquartile range (IQR), and whiskers extend to ±1.5 time the IQR; outliers are shown as black dots (pairwise comparisons using t-test with FDR correction, ***p.adj < 2e-16; n.s.: not significant). Bottom panel: amount of poly(A) RNA isolated from each organ by eRIC. Note that the retrieved mass of protein (upper panel) differs extensively across the tissues analyzed (with kidney >> liver > brain) and does not necessarily correlate with the mass of RNA recovered (bottom panel) (see also Table S2). (C) Hierarchical clustering and heatmap of the RBPs identified in brain, kidney and liver, showing protein abundance in eRIC eluates (left columns) and inputs (right columns) across the three organs. (D) Representative images of the proximity ligation assay (PLA) for interactions of ENO1 and poly(A) RNA, nuclear staining (DAPI) and ENO1 immunofluorescence in brain, kidney and liver. Scale bar, 20 µM. See quantification in Figure S2B.

Article Snippet: Subsequently, the Duolink PLA Fluorescence protocol (Sigma) was followed using an anti-biotin antibody (mouse: 1:400; ab201341, Abcam) and either the anti-ENO1 (rabbit: 1:400; Proteintech, 11204-1-AP), anti-SLC3A2 (rabbit: 1:400; Santa Cruz Biotechnology, sc-9160), anti-DDX6 (rabbit: 1:400; Novus Biologicals, NB200-192) or anti-PKM1 antibody (rabbit: 1:400; Cell Signalling, D30G6) for detection of the protein–RNA signal.

Techniques: Ex Vivo, Isolation, Proximity Ligation Assay, Staining, Immunofluorescence

Journal: Oncology Letters

Article Title: α-enolase is highly expressed in liver cancer and promotes cancer cell invasion and metastasis

doi: 10.3892/ol.2020.12003

Figure Lengend Snippet: ENO1 expression in liver cancer tissue and peripheral blood of patients with liver cancer. (A) ENO1 expression in liver cancer tissues (×100). (B) ENO1 was not expressed in benign liver lesions (×100). (C) Serum anti-ENO1 antibody levels in patients with liver cancer were higher than those in patients with benign liver lesions and healthy controls. (D) ENO1 expression in liver cancer tissues (×400). (E) ENO1 was not expressed in benign liver lesion (×400). (F) Receiver operating characteristic curve analysis for liver cancer diagnosis using anti-ENO1 antibody levels. The AUC was 0.741, the sensitivity was 64.3% and the specificity was 85.5%. The red dot represents the cut-off value. **P<0.01; ****P<0.0001 ENO1, α-enolase; AUC, area under the curve.

Article Snippet: The anti-ENO1 antibody ELISA detection reagent kit (cat. no. CSB-EQ027775HU) was purchased from CUSABIO Technology LLC.

Techniques: Expressing

Journal: Oncology Letters

Article Title: α-enolase is highly expressed in liver cancer and promotes cancer cell invasion and metastasis

doi: 10.3892/ol.2020.12003

Figure Lengend Snippet: ENO1 expression in pathological tissues.

Article Snippet: The anti-ENO1 antibody ELISA detection reagent kit (cat. no. CSB-EQ027775HU) was purchased from CUSABIO Technology LLC.

Techniques: Expressing

Journal: Oncology Letters

Article Title: α-enolase is highly expressed in liver cancer and promotes cancer cell invasion and metastasis

doi: 10.3892/ol.2020.12003

Figure Lengend Snippet: Comparison of the serum anti-ENO1 antibody levels among the three groups of participants [P50 (P25-P75)].

Article Snippet: The anti-ENO1 antibody ELISA detection reagent kit (cat. no. CSB-EQ027775HU) was purchased from CUSABIO Technology LLC.

Techniques:

Journal: Oncology Letters

Article Title: α-enolase is highly expressed in liver cancer and promotes cancer cell invasion and metastasis

doi: 10.3892/ol.2020.12003

Figure Lengend Snippet: Validation of the siRNA interference effect on ENO1 expression in liver cancer cells. (A) RT-qPCR validation of the effect of siRNA interference on ENO1 expression in HepG2 cells. The results showed that the relative expression levels of the ENO1 gene in the si-1 and si-2 groups were lower than those in the control group. (B) RT-qPCR validation of the effect of siRNA interference on ENO1 expression in Huh7 cells. The results showed that the relative expression levels of the ENO1 gene in the si-1 and si-2 groups were lower than those in the control group. (C) Western blot validation of the effect of siRNA interference on ENO1 expression in HepG2 cells. The results showed that the relative expression levels of the ENO1 protein in HepG2 cells in the si-1 and si-2 groups were significantly lower than those in the control group. (D) Western blot validation of the effect of siRNA interference on ENO1 expression in Huh7 cells. The results showed that the relative expression levels of the ENO1 protein in Huh7 cells in the si-1 and si-2 groups were significantly lower than those in the control group. **P<0.01; ****P<0.0001. siRNA, small interfering RNA; ENO1, α-enolase; si-1, siRNA interference group 1; si-2, siRNA interference group 2; RT-qPCR, reverse transcription-quantitative PCR.

Article Snippet: The anti-ENO1 antibody ELISA detection reagent kit (cat. no. CSB-EQ027775HU) was purchased from CUSABIO Technology LLC.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Small Interfering RNA, Real-time Polymerase Chain Reaction

Journal: Oncology Letters

Article Title: α-enolase is highly expressed in liver cancer and promotes cancer cell invasion and metastasis

doi: 10.3892/ol.2020.12003

Figure Lengend Snippet: Effect of ENO1 siRNA on liver cancer cell proliferation. (A) HepG2 and (B) Huh7 cell proliferation was suppressed after ENO1 siRNA treatment. Results show that 72 h after transfection in the si-1 and si-2 groups, ENO1 siRNA treatment resulted in proliferation inhibition in HepG2 and Huh7 cells. In HepG2 cells, the differences in cell proliferation between the NC and the siRNA-treated groups were statistically significant (P<0.05 vs. si-2 group and P<0.01 vs. si-1 group). Cell proliferation in Huh7 cells was also significantly different. *P<0.05; **P<0.01. ENO1, α-enolase; siRNA, small interfering RNA; si-1, siRNA interference group 1; si-2, siRNA interference group 2; OD, optical density; NC, negative control.

Article Snippet: The anti-ENO1 antibody ELISA detection reagent kit (cat. no. CSB-EQ027775HU) was purchased from CUSABIO Technology LLC.

Techniques: Transfection, Inhibition, Small Interfering RNA, Negative Control

Journal: Oncology Letters

Article Title: α-enolase is highly expressed in liver cancer and promotes cancer cell invasion and metastasis

doi: 10.3892/ol.2020.12003

Figure Lengend Snippet: Effect of ENO1 siRNA on the migration ability of liver cancer cells. (A) Compared with that of the NC group, HepG2 cell migration after ENO1 siRNA treatment became slower in the si-1 and si-2 groups, and the difference was statistically significant at 48 h after treatment. (B) Gap size was measured using ImageJ software and the ratio of gap size after ENO1 siRNA treatment in HepG2 cells was calculated based on the size of the wound at 0 h. (C) Compared with that of the NC group, Huh7 cell migration after ENO1 siRNA treatment became slower in the si-1 and si-2 groups, and the difference was statistically significant at 48 h after treatment. (D) Gap size was measured with the ImageJ software and the ratio of gap size after ENO1 siRNA treatment in Huh7 cell was calculated based on the size of the wound at 0 h. *P<0.05; **P<0.01. ENO1, α-enolase; siRNA, small interfering RNA; NC, negative control; si-1, siRNA interference group 1; si-2, siRNA interference group 2.

Article Snippet: The anti-ENO1 antibody ELISA detection reagent kit (cat. no. CSB-EQ027775HU) was purchased from CUSABIO Technology LLC.

Techniques: Migration, Software, Small Interfering RNA, Negative Control

Journal: Oncology Letters

Article Title: α-enolase is highly expressed in liver cancer and promotes cancer cell invasion and metastasis

doi: 10.3892/ol.2020.12003

Figure Lengend Snippet: Effect of ENO1 siRNA on the invasion and migration abilities of liver cancer cells. Compared with the NC group, the in vitro invasion and migration abilities of (A) HepG2 and (B) Huh7 cells after ENO1 siRNA treatment decreased in the si-1 and si-2 groups, and the differences were statistically significant. Numbers of (C) HepG2 and (D) Huh7 cells were measured with the ImageJ software after ENO1 siRNA treatment. *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001. ENO1, α-enolase; siRNA, small interfering RNA; NC, negative control; si-1, siRNA interference group 1; si-2, siRNA interference group 2.

Article Snippet: The anti-ENO1 antibody ELISA detection reagent kit (cat. no. CSB-EQ027775HU) was purchased from CUSABIO Technology LLC.

Techniques: Migration, In Vitro, Software, Small Interfering RNA, Negative Control