Extended Data Fig. 2: Characterization of spontaneously metastatic orthotopic lung LLC and subcutaneous EO771 – LMB models.
From: Radiation-induced amphiregulin drives tumour metastasis
a, Areg upregulation in LLCAR+ and LLCAR− cells after 4 Gy RT in vitro, measured by RT-qPCR 3 h post-RT. One-way ANOVA, Šídák’s test. n = 3 technical repeats. b, AREG concentration in flank tumour lysate ± 20 Gy RT, measured by ELISA 5 days post-RT, n = 5 mice in AR+ groups, n = 4 in AR- groups. One-way ANOVA, Šídák’s test. c, Serum AREG concentration in tumour-bearing mice ± 20 Gy RT, measured by ELISA. Blood draw timepoints post-RT as indicated, ctrl - n = 3 non-tumour bearing mice, n = 5 mice in other groups. Two-way ANOVA, Tukey’s test. d, CT-images of orthotopic LLCAR+ and LLCAR− tumours on days 8, 11, and 14 post-injection. White asterisks/red highlights indicate tumours. e, Growth curves of LLCAR+ and LLCAR− orthotopic lung tumours ± 8 or 12 Gy CT-guided RT. n = 6 mice per group. Two-way ANOVA, Šídák’s test. f, Serum AREG concentration of LLCAR+ and LLCAR− orthotopic lung tumour-bearing mice five days after 8 or 12 Gy RT, measured by ELISA, n = 4 mice in all groups. One-way ANOVA, Šídák’s test. g, Number of contralateral lung metastases (left) and mean size of the five largest metastases per contralateral lung area (right) from LLCAR+ and LLCAR− orthotopic tumours (n = 4 in LLCAR+, n = 5 in LLCAR−, n = 6 mice in others). One-way ANOVA, Šídák’s test. h, H&E staining of LLCAR+ and LLCAR− orthotopic lung tumours. Black arrows indicate contralateral intrapulmonary metastatic lesions, white asterisks = orthotopic tumours, LN = extrapulmonary, intrathoracic lymph node metastasis. i, AREG MFI in LMB and LLC tumour cells post 4 Gy and 8 Gy RT in vitro, determined by flow cytometry. n = 2 technical repeats in LLC + 8 Gy, n = 3 in all other groups. j, Growth curves of LMB flank tumours ± RT with indicated doses, n = 4 mice in all groups, two independent experiments. Two-way ANOVA, Tukey’s test. k, Number of LMB lung metastases (left) and mean size of the five largest lung metastases per lung area (right), n = 5 mice in LMB and 5 Gy groups, n = 4 in 10 Gy and 20 Gy groups. One-way ANOVA, Dunnett’s test. l, H&E staining. Black arrows indicate metastatic lesions. RT doses as indicated. m, Areg upregulation in LMBAR+ and LMBAR− cells after 4 Gy RT in vitro post-RT, RT-qPCR. One-way ANOVA, Šídák’s test. n = 3 technical repeats. n, Growth curves of LMBAR+ and LMBAR− flank tumours ± 5 Gy; n = 4 mice in LMBAR− + 5 Gy group, n = 5 mice in all others. o, Number of lung metastases (left), and mean size of five largest metastases per lung area (right), n = 4 mice in AR+ groups, n = 5 in AR- groups. One-way ANOVA, Šídák’s test. p, H&E staining. Black arrows indicate metastatic lesions. Data are shown as mean ± SEM; box plots show median (Q2), Q1–Q3, and whiskers (min–max).
