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. 2022 Sep 6;11(18):2783. doi: 10.3390/cells11182783

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© 2022 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Immune phenotyping of JQ1-treated TH-MYCN tumors. (A) Experimental design of JQ1 treatment of TH-MYCN tumor-bearing mice showing the schedule and dosing. Following the development of homozygote TH-MYCN tumors, the tumor volume was assessed by MRI in all mice. Mice were randomly assigned to a vehicle-treated group (n = 6) or a JQ1-treated group (n = 5). Mice were treated three times with vehicle or JQ1 (25 mg/kg) by IP on days 1, 2, and 3. On day 4 post-treatment, MRI was performed to determine the tumor volume, and tumors were harvested for immune phenotyping experiments. (B–D) Flow cytometry quantification of total (B), CD69⁺ (C), or PD-1⁺ (D) CD8⁺ T cells, conventional (Conv.) CD4⁺ T cells and Tregs infiltrating vehicle-treated or JQ-1-treated TH-MYCN tumors at day 4. The defined subpopulations were gated and quantified in live CD45⁺ cells. Each dot represents one tumor. The data are reported as the average of six or five mice per group. Results are shown as mean ± SEM (error bars). Statistically significant differences (indicated by asterisks) are calculated compared to vehicle-treated tumors using an unpaired two-tailed Student’s t-test (ns: not significant *: p < 0.05).

Immune phenotyping of JQ1-treated TH-MYCN tumors. (A) Experimental design of JQ1 treatment of TH-MYCN tumor-bearing mice showing the schedule and dosing. Following the development of homozygote TH-MYCN tumors, the tumor volume was assessed by MRI in all mice. Mice were randomly assigned to a vehicle-treated group (n = 6) or a JQ1-treated group (n = 5). Mice were treated three times with vehicle or JQ1 (25 mg/kg) by IP on days 1, 2, and 3. On day 4 post-treatment, MRI was performed to determine the tumor volume, and tumors were harvested for immune phenotyping experiments. (B–D) Flow cytometry quantification of total (B), CD69⁺ (C), or PD-1⁺ (D) CD8⁺ T cells, conventional (Conv.) CD4⁺ T cells and Tregs infiltrating vehicle-treated or JQ-1-treated TH-MYCN tumors at day 4. The defined subpopulations were gated and quantified in live CD45⁺ cells. Each dot represents one tumor. The data are reported as the average of six or five mice per group. Results are shown as mean ± SEM (error bars). Statistically significant differences (indicated by asterisks) are calculated compared to vehicle-treated tumors using an unpaired two-tailed Student’s t-test (ns: not significant *: p < 0.05).