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Transcriptomic analysis reveals temporal responses to E2 and HDACi. RNA‐seq was performed using HCC‐1428 and HCC‐1428/LTED cells in HD medium treated in triplicate as described in (A). (B, C) Heatmap of the 500 most variably expressed genes across all treatment groups within each cell line. (D) Volcano plots of differentially expressed genes after 1 day of drug treatments. Red dots indicate significantly altered genes ( P ≤ 0.05) with |log 2 fold‐change| ≥ 1.

Journal: Molecular Oncology

Article Title: Pharmacological chromatin remodeling enhances response to estrogen therapy in ER + breast cancer

doi: 10.1002/1878-0261.70307

Figure Lengend Snippet: Transcriptomic analysis reveals temporal responses to E2 and HDACi. RNA‐seq was performed using HCC‐1428 and HCC‐1428/LTED cells in HD medium treated in triplicate as described in (A). (B, C) Heatmap of the 500 most variably expressed genes across all treatment groups within each cell line. (D) Volcano plots of differentially expressed genes after 1 day of drug treatments. Red dots indicate significantly altered genes ( P ≤ 0.05) with |log 2 fold‐change| ≥ 1.

Article Snippet: For tumor RNA‐seq analysis, extracted RNA was used for RNA sequencing by Plasmidsaurus using Illumina Sequencing Technology.

Techniques: RNA Sequencing

E2 and HDACi synergistically induce tumor regression and prevent recurrence. (A, B) Ovx mice bearing CTG‐3346 or WHIM16 tumors were randomized to drug treatments; animal numbers are indicated in keys. E2 was delivered continuously, and entinostat was administered 5 days·week −1 for 7.5 weeks (gray shading). Tumor growth curves (A) are shown as mean + SEM. **** P < 0.001 by linear mixed modeling of raw tumor volumes. After 7.5 weeks of E2 ± entinostat treatment, mice with completely regressed tumors were monitored for recurrence in (B), defined as regrowth to 200 mm 3 . (C, D) Tumors were harvested 4 h after drug treatment on Day 3 or 14. Tumor lysates ( n = 3/group) were analyzed by immunoblot. Mean (SD) band intensities for each treatment group are noted below each blot. Cleaved PARP was normalized to full‐length PARP prior to normalization to vinculin loading control. (E) Tumors from (C/D) were analyzed by RNA‐seq. Transcriptomic profiles were analyzed by GSEA using Hallmark pathways. Dot color reflects NES for comparison of each treatment group vs. vehicle, or the drug combination vs. E2 as indicated.

Journal: Molecular Oncology

Article Title: Pharmacological chromatin remodeling enhances response to estrogen therapy in ER + breast cancer

doi: 10.1002/1878-0261.70307

Figure Lengend Snippet: E2 and HDACi synergistically induce tumor regression and prevent recurrence. (A, B) Ovx mice bearing CTG‐3346 or WHIM16 tumors were randomized to drug treatments; animal numbers are indicated in keys. E2 was delivered continuously, and entinostat was administered 5 days·week −1 for 7.5 weeks (gray shading). Tumor growth curves (A) are shown as mean + SEM. **** P < 0.001 by linear mixed modeling of raw tumor volumes. After 7.5 weeks of E2 ± entinostat treatment, mice with completely regressed tumors were monitored for recurrence in (B), defined as regrowth to 200 mm 3 . (C, D) Tumors were harvested 4 h after drug treatment on Day 3 or 14. Tumor lysates ( n = 3/group) were analyzed by immunoblot. Mean (SD) band intensities for each treatment group are noted below each blot. Cleaved PARP was normalized to full‐length PARP prior to normalization to vinculin loading control. (E) Tumors from (C/D) were analyzed by RNA‐seq. Transcriptomic profiles were analyzed by GSEA using Hallmark pathways. Dot color reflects NES for comparison of each treatment group vs. vehicle, or the drug combination vs. E2 as indicated.

Article Snippet: For tumor RNA‐seq analysis, extracted RNA was used for RNA sequencing by Plasmidsaurus using Illumina Sequencing Technology.

Techniques: Western Blot, Control, RNA Sequencing, Comparison