Fig. 7: In vivo effect of ANP32E overexpression and ATRi treatment. | Nature Communications

Fig. 7: In vivo effect of ANP32E overexpression and ATRi treatment.

From: ANP32E drives vulnerability to ATR inhibitors by inducing R-loops-dependent transcription replication conflicts in triple negative breast cancer

Fig. 7

a Representative PLA images showing TRCs between pS2RNApol II and PCNA in mouse xenografts derived from tIMEC and tIMEC-A cells. Three zoom-ins of single cells are shown per condition. Scale bar = 10 µm. b Representative immunofluorescence images of pRPA32 (red) and human-mitochondria (green) in xenografts from tIMEC and tIMEC-A. Three zoom-ins per condition are shown. c Violin plots showing the number of PLA (left) or pRPA32 (right) foci per nucleus across two xenografts per condition, corresponding to (a, b). Unpaired two-tailed t-test p values are indicated. d Dose-response curves of tIMEC, tIMEC-A, and tIMEC-A-H1 cells treated with VE822 for 48 h. EC50 indicated by vertical dashed lines. Data were reported as Mean ± SD of three technical replicates. e Experimental scheme: luciferase-tagged tIMEC or tIMEC-A cells were orthotopically injected into mice. Once tumors were palpable, VE822 (4x/week) or vehicle was administered. After 4 weeks, tumors and metastases were analyzed. f Dot plot representing tumor growth curves (mean ± s.e.m.) for tIMEC and tIMEC-A xenografts with or without VE822 treatment, monitored over 21 days. Two-way ANOVA p values are shown. n: tIMEC vehicle = 11, VE822 = 9; tIMEC-A vehicle = 10, VE822 = 11 (pooled from two independent experiments). g Barplot showing mean ± s.e.m. photon flux per metastasis in vehicle- or VE822-treated mice with tIMEC or tIMEC-A tumors. n: tIMEC vehicle = 4, VE822 = 3; tIMEC-A vehicle = 5, VE822 = 4. Two-tailed t-test p values indicated. Representative lung metastases are shown below. h Mean cell number per metastasis (±SD) in tIMEC or tIMEC-A tumors ± VE822. n: tIMEC vehicle = 8, VE822 = 3; tIMEC-A vehicle = 27, VE822 = 43. Two-tailed unpaired t-test p values are shown. i Left: Barplot of cell number per metastasis (mean ± s.e.m.) categorized into DTCs, micro-, and macrometastases. Two-tailed Wilcoxon t-test p values are shown. Right: Representative IHC images (human-mitochondria) identifying metastasis types. Scale bar = 100 µm; zoom-in = 15 µm. Cell counts used to classify each category are provided.

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