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. 2020 Oct 20;33(3):108286. doi: 10.1016/j.celrep.2020.108286

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Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.

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Senescence and Senescence-Associated Gene Expressions and Secretory Phenotype (SASP) with BMI1 Inhibition and Impact of BH3 Mimetics on BMI1-Inhibition-Induced Cellular Senescence and Apoptosis

(A and B) Representative pictures of SA-β-gal (senescence associated beta galactosidase) staining with genetic inhibition of BMI1 (A) and with BMI1 inhibitors (B) in SF8628. Adjacent graphs show the quantification of the number of cells with blue staining normalized to the total number of cells. By ANOVA, the p value within the three groups was p < 0.0001 (A) and p = 0.0037 (B).

(C) Western blot analysis of p16 and p21 in SU-DIPG04 cells transduced with shBMI1.

(D) Western blot analysis of p16 and p21 in SF8628 cells treated with PTC028.

(E–G) Concentration of SASP factors released by DIPG cells after treatment with either PTC209 (E) or PTC028 (F) and after genetic knockdown of BMI1 (G).

(H) Changes in BMI1 and H2Aub genome tracks at the specific gene locations indicated in SU-DIPG04 cells after PTC028 treatment. The x axis represents genome locations, and the y axis represents the RPM for the indicated genes. For RNA-seq tracks, the increase in gene expression with PTC028 treatment is visualized by greater transcript tag density over the exons of the indicated genes (exons are the horizontal bars, and the direction of transcription is indicated by arrows shown below ChIP-seq tracks).

(I) Representative pictures of neurospheres and sub-spheres (red arrows) formed in short- and long-term exposure of DIPG cells to PTC028, with quantification of size and number of neurospheres formed shown adjacently.

(J) SASP factors measured in SU-DIPG04 with long-term exposure to PTC209.

(K) Western blot showing changes in anti-apoptotic proteins after short- and long-term exposure of cells to PTC drugs.

(L) Dose-response curve on cell viability (MTS (viability asassay endpoint) of SF8628 cells treated with PTC028 and ABT263 either alone or in combination. MTS assay is a viability assay based on cell metabolic activity.

(M) Real-time measurement of proliferation of HSJD-DIPG-007 shBMI1 or shNull cells with ABT263 (250 nM).

(N) Representative pictures of SA-β-gal staining of BT245 cells, with different treatment conditions shown.

(O) Flow cytometry analysis of annexin V-PI/7AAD as a measure of apoptosis in HSJD-DIPG-007 cells treated with PTC028 and obatoclax. By ANOVA, ^∗∗∗∗p < 0.0001. Pairwise comparisons; ^∗p < 0.05, ^∗∗p < 0.012, ^∗∗∗∗p < 0.001; control versus treatment by Student’s t test. Data represent mean ± SEM.

See also Figures S7 and S8.