Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2019 May 13;42(1):55–66. doi: 10.3892/or.2019.7156

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

Copyright: © Zhang et al.

This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.

PMC Copyright notice

Figure 4. — SUZ12 reverses the inhibitory effects of miR-767-5p on GBM cell lines. (A) The expression of SUZ12 in U87 and U251 cells was calculated by RT-qPCR after transfection. (B-I) For all assays, U87 and U251 cells were transfected with miR-767-5p mimic, negative control sequence (miR-NC), or miR-767-5p and a SUZ12 overexpression plasmid. (B) CCK-8 cell proliferation assay. (C and D) Representative images (left) and quantification (right) of (C) colony-forming assays, (D) wound healing assays, and (E) Transwell invasion assays. (F) Flow cytometric analysis of the cell cycle analyzed 48 h after transfection representative histograms (left) and quantification (right) of the percentage of cells in G1, G2, and S phases of the cell cycle. (G) Representative dot plots (left) and quantification (right) of an Annexin V-FITC/PI apoptosis assay. (H) Representative images of immunofluorescence staining of SUZ12 in U87 and U251 cells. Nuclei were stained with DAPI. (H) Representative images of immunofluorescence staining of SUZ12 in U87 and U251 cells. Nuclei were stained with DAPI. (I) Western blot analysis of SUZ12, phosphorylated (p)-ERK1/2, total ERK1/2, p-AKT, and total AKT levels (left) and quantification of relative p-AKT and p-ERK levels (right). β-actin was probed as an internal control. Data are presented as the mean ± SD of n=3. *P<0.05, **P<0.01, miR-767-5p vs. miR-767-5p+SUZ12. SUZ12, suppressor of zeste-12; GBM, glioblastoma multiforme; CCK-8, Cell Counting Kit-8.

Figure 4. — SUZ12 reverses the inhibitory effects of miR-767-5p on GBM cell lines. (A) The expression of SUZ12 in U87 and U251 cells was calculated by RT-qPCR after transfection. (B-I) For all assays, U87 and U251 cells were transfected with miR-767-5p mimic, negative control sequence (miR-NC), or miR-767-5p and a SUZ12 overexpression plasmid. (B) CCK-8 cell proliferation assay. (C and D) Representative images (left) and quantification (right) of (C) colony-forming assays, (D) wound healing assays, and (E) Transwell invasion assays. (F) Flow cytometric analysis of the cell cycle analyzed 48 h after transfection representative histograms (left) and quantification (right) of the percentage of cells in G1, G2, and S phases of the cell cycle. (G) Representative dot plots (left) and quantification (right) of an Annexin V-FITC/PI apoptosis assay. (H) Representative images of immunofluorescence staining of SUZ12 in U87 and U251 cells. Nuclei were stained with DAPI. (H) Representative images of immunofluorescence staining of SUZ12 in U87 and U251 cells. Nuclei were stained with DAPI. (I) Western blot analysis of SUZ12, phosphorylated (p)-ERK1/2, total ERK1/2, p-AKT, and total AKT levels (left) and quantification of relative p-AKT and p-ERK levels (right). β-actin was probed as an internal control. Data are presented as the mean ± SD of n=3. *P<0.05, **P<0.01, miR-767-5p vs. miR-767-5p+SUZ12. SUZ12, suppressor of zeste-12; GBM, glioblastoma multiforme; CCK-8, Cell Counting Kit-8.