Journal: Oncology reports

Article Title: miRNA-regulated expression of oncogenes and tumor suppressor genes in the cisplatin-inhibited growth of K562 cells.

doi: 10.3892/or_00000813

Figure Lengend Snippet: Figure 2. Changes in mRNA expression of BCL2, E2F1, E2F3, RB1 and P53 in K562 cells after cisplatin treatment. BCL2, E2F1, E2F3, RB1 and P53 were detected by (A) RT-PCR, (B) real-time PCR and (C) ELISA.

Article Snippet: K562 cells were lysed [lysis buffer: 0.15 M NaCl, 5 mM EDTA (pH 8.0), 1% Triton X-100, 10 mM Tris-Cl (pH 7.4), 100 mM PMSF and 5 M DTT) and incubated in a 96-well plate, followed by the addition of goat anti-human antibodies against BCL2 (1:400), E2F1, E2F3 (1:400, 1:400), RB1 (1:500) and P53 (1:300, 1:400) (all from Immunoleader, Boster) according to the manufacturer's instructions.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: Oncology reports

Article Title: miRNA-regulated expression of oncogenes and tumor suppressor genes in the cisplatin-inhibited growth of K562 cells.

doi: 10.3892/or_00000813

Figure Lengend Snippet: Figure 4. Correlative expression of miRNAs and oncogenes using antisense oligos (ASO). (A) RT-PCR, (B) real-time PCR. Correlative expression of (C) E2F1 and its targeted miR-17-5p, (D) E2F3 and its targeted miRNAs and (E) Bcl-2 and its targeted miRNAs (miR-16, 34a-c) using ELISA is shown. *Significant difference (p<0.05).

Article Snippet: K562 cells were lysed [lysis buffer: 0.15 M NaCl, 5 mM EDTA (pH 8.0), 1% Triton X-100, 10 mM Tris-Cl (pH 7.4), 100 mM PMSF and 5 M DTT) and incubated in a 96-well plate, followed by the addition of goat anti-human antibodies against BCL2 (1:400), E2F1, E2F3 (1:400, 1:400), RB1 (1:500) and P53 (1:300, 1:400) (all from Immunoleader, Boster) according to the manufacturer's instructions.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: Oncology reports

Article Title: miRNA-regulated expression of oncogenes and tumor suppressor genes in the cisplatin-inhibited growth of K562 cells.

doi: 10.3892/or_00000813

Figure Lengend Snippet: Figure 5. Expression of E2F1, E2F3, BCL2, RB1 and P53 genes regulated by miRNAs. Detection of the expression of (A) E2F1, (B) E2F3, (C) BCL2, (D) RB1 and (E) P53. *Significant difference (p<0.05).

Article Snippet: K562 cells were lysed [lysis buffer: 0.15 M NaCl, 5 mM EDTA (pH 8.0), 1% Triton X-100, 10 mM Tris-Cl (pH 7.4), 100 mM PMSF and 5 M DTT) and incubated in a 96-well plate, followed by the addition of goat anti-human antibodies against BCL2 (1:400), E2F1, E2F3 (1:400, 1:400), RB1 (1:500) and P53 (1:300, 1:400) (all from Immunoleader, Boster) according to the manufacturer's instructions.

Techniques: Expressing

Journal: International journal of biological sciences

Article Title: Integrative analysis of multi-omics data reveals inhibition of RB1 signaling promotes apatinib resistance of hepatocellular carcinoma.

doi: 10.7150/ijbs.83862

Figure Lengend Snippet: Figure 4. Changes in chromatin accessibility are consistent with global transcriptome changes. A. Genome tracks showing a comparison of ATAC-seq profiles of RB1 in apatinib-resistant and non-resistant HepG2 cells. B. The mRNA expression levels of RB1 in apatinib-resistant versus untreated HepG2 cells, statistical analysis was performed by Student t-test. C. Boxplots of chromatin accessibility values for high expression genes and low expression genes in apatinib-resistant cells.

Article Snippet: After overnight incubation with primary antibody (anti - RB1 and CDKN2B antibody; Boster Bio, Pleasanton, USA) by 1:200 at 4 °C, sections were incubated for 15 minutes at room temperature with horseradish peroxidaselabeled polymer conjugated with secondary antibody (MaxVision Kits) and incubated for 1 minute with diaminobenzidine.

Techniques: Comparison, Expressing

Journal: International journal of biological sciences

Article Title: Integrative analysis of multi-omics data reveals inhibition of RB1 signaling promotes apatinib resistance of hepatocellular carcinoma.

doi: 10.7150/ijbs.83862

Figure Lengend Snippet: Figure 6. Expression status of RB1 pathway targets in HCC cells and in clinical samples; CDK2-IN-73 and Palbociclib can relieve apatinib resistance. A. qPCR analysis showed that the expression levels of RB1, PCNA, KIAA0101 and MCM7 in HepG2-resistant cells were significantly reduced, and the expression levels of CDKN2B andCDKN2AIP were increased. B. Western blot detection of the above proteins is consistent with qPCR detection results. C. Analysis of mRNA expression of five PD patients showed high expression of CDKN2B and low expression of RB1, PCNA, and MCM7. D. Western blot analysis showed that PD patients had high expression of CDKN2B but low expression of RB1, PCNA, and MCM7. E. Representative photographs of staining of RB1 protein in a pair of HCC patients tissues in PD and PR (20X). F. Representative photographs of staining of CDKN2B protein in a pair of HCC patients tissues in PD and PR (20X). G. The proliferation inhibition curves of resistant HepG2 cells and HepG2 cells treated with CDK2-IN-73. H. Western blot analysis showed that the expression of CDKN2B in HepG2resistant cells was not significantly different, and the expression levels of RB1, PCNA and MCM7 were upregulated. I. Proliferation inhibition curves of HepG2 resistant cells and HepG2 cells treated with Palbociclib. J. Western blot analysis results showed that the expression levels of RB1, PCNA, and MCM7 were upregulated. The bars represent mean ± standard deviation; ∗P < 0.05.

Article Snippet: After overnight incubation with primary antibody (anti - RB1 and CDKN2B antibody; Boster Bio, Pleasanton, USA) by 1:200 at 4 °C, sections were incubated for 15 minutes at room temperature with horseradish peroxidaselabeled polymer conjugated with secondary antibody (MaxVision Kits) and incubated for 1 minute with diaminobenzidine.

Techniques: Expressing, Western Blot, Staining, Inhibition, Standard Deviation