brd2  (Bethyl)

Journal: Cell death & disease

Article Title: PROTAC induced-BET protein degradation exhibits potent anti-osteosarcoma activity by triggering apoptosis.

doi: 10.1038/s41419-019-2022-2

Figure Lengend Snippet: Fig. 2 BETd-260 potently induces degradation of BET proteins in OS cells. a MNNG/HOS and b Saos-2 cell lines were treated with BETd-260, HJB-97, or JQ1 as indicated for 24 h. The protein levels of BRD2, 3, and 4 were examined by western blotting analysis. Tubulin was used as a loading control. c The MNNG/HOS cell line was treated with BETd-260 at 30 nM for different times. The protein level of BRD2, 3, and 4 were examined by western blotting analysis. Tubulin was used as a loading control. d MNNG/HOS cells were pretreated with MG-132 (2000 nmol/L), MNL4924 (500 nmol/L), or HJB-97 (3000 nmol/L) for 1 h, followed by treatment with BETd-260 (10 nM) for 2 h. The protein levels of BRD2, 3, and 4 were examined by western blotting analysis. Tubulin was used as a loading control. Data are representative of three independent experiments

Article Snippet: The following antibodies were used for IHC: BRD2 (A302-583A, 1:250), BRD3 (A302-368A, 1:250), BRD4 (A700-005, 1:100) from Bethyl Laboratories (Shanghai, China); cleaved PARP-1 (Asp214) (5625, 1:100) and Ki67 (8D5) (9449, 1:500) from Cell Signaling Technology (CST, Shanghai, China).

Techniques: Western Blot, Control

Journal: Cell death & disease

Article Title: PROTAC induced-BET protein degradation exhibits potent anti-osteosarcoma activity by triggering apoptosis.

doi: 10.1038/s41419-019-2022-2

Figure Lengend Snippet: Fig. 5 BETd-260 induces BET degradation, triggers apoptosis in vivo, and inhibits tumor growth in MNNG/HOS xenografts in mice. a, b BALB/c mice bearing MNNG/HOS xenograft tumors were treated with a single intravenous dose of 5 mg/kg BETd-260. Two to three mice were sacrificed and tumor tissue was harvested at each different time points. a The levels of BRD2, BRD3, and BRD4 proteins in tumor tissue lysates were examined by western blotting analysis. Tubulin was used as a loading control. b The expression of BRD2, BRD3, BRD4, cleaved PARP-1, and Ki67 was examined by immunohistochemistry staining. c–e BALB/c mice bearing xenograft tumors were treated with BETd-260 and vehicle control intravenously three times per week for 3 weeks. Tumor sizes and body weights were measured every 2–3 days. Data are mean ± SEM (n = 7–8). c Tumor values were plotted with Prism 6 software. d Scatter plots represent the tumor volumes at the end of each study. e Body weights were plotted with Prism 6 software. p values between each treated and the vehicle control group were determined using two-way ANOVA. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001)

Article Snippet: The following antibodies were used for IHC: BRD2 (A302-583A, 1:250), BRD3 (A302-368A, 1:250), BRD4 (A700-005, 1:100) from Bethyl Laboratories (Shanghai, China); cleaved PARP-1 (Asp214) (5625, 1:100) and Ki67 (8D5) (9449, 1:500) from Cell Signaling Technology (CST, Shanghai, China).

Techniques: In Vivo, Western Blot, Control, Expressing, Immunohistochemistry, Staining, Software

Journal: Cell death & disease

Article Title: PROTAC induced-BET protein degradation exhibits potent anti-osteosarcoma activity by triggering apoptosis.

doi: 10.1038/s41419-019-2022-2

Figure Lengend Snippet: Fig. 6 BETd-260 induces BET degradation, triggers apoptosis in vivo, and inhibits tumor growth in PDX xenografts in mice. a NOD SCID mice bearing PDX xenograft tumors were treated with a single intravenous dose of 5 mg/kg BETd-260. Two to three mice were sacrificed and tumor tissue was harvested at each different time points. The expression of BRD2, BRD3, BRD4, cleaved PARP-1, and Ki67 was examined by immunohistochemistry staining. b–d NOD SCID mice bearing xenograft tumors were treated with BETd-260 or vehicle control intravenously three times per week for 3 weeks. Tumor sizes and body weights were measured every 2–3 days. Data are mean ± SEM (n = 7–8). b Tumor values were plotted with Prism 6 software. c Scatter plots represent the tumor volumes at the end of each study. d Body weights were plotted with Prism 6 software. p values between each treated and the vehicle control group were determined using two-way ANOVA. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001

Article Snippet: The following antibodies were used for IHC: BRD2 (A302-583A, 1:250), BRD3 (A302-368A, 1:250), BRD4 (A700-005, 1:100) from Bethyl Laboratories (Shanghai, China); cleaved PARP-1 (Asp214) (5625, 1:100) and Ki67 (8D5) (9449, 1:500) from Cell Signaling Technology (CST, Shanghai, China).

Techniques: In Vivo, Expressing, Immunohistochemistry, Staining, Control, Software