Supplementary Information s10

SUPPLEMENTARY INFORMATION

Annals of Surgical Oncology

Elevated expression of Girdin in the nucleus indicates worse prognosis for patients with estrogen

receptor-positive breast cancer

Wen-Ting Peng* M.D.; Xin Hu* Ph.D.; Ling Yao* Ph.D.; Yi-Zhou Jiang M.D.; Zhi-Ming Shao# M.D.

* These authors contributed equally to this work.

Correspondence should be addressed to Zhi-Ming Shao M.D.

Department of Breast Surgery

Shanghai Cancer Center and Cancer Institute

Fudan University

399 Ling-Ling Road, Shanghai, 200032, P.R. China

Email: [email protected] Supplementary Table 1 Correlations between patient characteristics and the expression status of phospho-

Akt (S473) and phospho-Akt (T308) in breast cancer

Clinical-pathological phospho-Akt (S473) phospho-Akt (T308) Cases Cases characteristics - + Pa - + Pa 250 50 200 248 72 176 Percentage (%) 20.00 80.00 29.03 70.97 Age (years) ≤50 120 26 94 0.527 119 38 81 0.334 >50 130 24 106 129 34 95 Menopausal status Pre 108 23 85 0.655 108 33 75 0.643 Post 142 27 115 140 39 101 TNM Stage I 74 14 60 0.720 74 19 55 0.540 II 132 29 103 130 42 88 III 42 7 35 42 11 31 Histological grade I 5 1 4 0.586 5 1 4 0.840 II 184 34 150 182 52 130 III 61 15 46 61 19 42 Tumor size T1 (≤ 2cm) 115 24 91 0.714 113 33 80 0.917 T2 (> 2-5cm) 120 25 95 120 35 85 T3 (> 5cm) 13 1 12 13 4 9 Node status Negative 151 31 120 0.857 151 40 111 0.301 Positive 97 19 78 95 31 64 ER status Negative 143 35 108 0.044 142 45 97 0.307 Positive 106 15 91 105 27 78 PR status Negative 184 38 146 0.583 183 54 129 0.943 Positive 63 11 52 62 18 44 HER-2 status Negative 148 34 114 0.168 148 55 93 0.001 Positive 101 16 85 99 17 82 Local recurrence Negative 175 40 135 0.076 174 51 123 0.511 Positive 26 2 24 26 6 20 Distant metastasis Negative 152 37 115 0.034 151 44 107 0.725 Positive 49 5 44 49 13 36 Abbreviations: ER, estrogen receptor; PR, progesterone receptor; HER-2, human epidermal growth factor receptor 2; a Pearson 2 test. Supplementary Figures

Fig. S1 Expression patterns of Girdin in the nucleus and cytoplasm, phospho-Akt (S473), and phospho- Akt (T308) in breast cancer. a Representative immunohistochemical (IHC) staining of negative and positive Girdin staining in the nucleus and cytoplasm. b Representative IHC staining of negative and positive phospho-Akt (S473) and phospho-Akt (T308) staining. For Fig. 1a, the scale bars represent 100

µm for the small pictures and 50 µm for the large ones; for Fig. 1b, the scale bars represent 100 µm for the small pictures and 20 µm for the large ones

Fig. S2 Cumulative disease-free survival (DFS) and overall survival (OS) curves of patients with positive or negative phospho-Akt (S473). a Positive phospho-Akt (S473) expression in breast cancer was associated with shortened DFS. b There were no significant differences in OS between phospho-Akt

(S473)-negative and phospho-Akt (S473)-positive breast cancer patients

Fig. S3 There were no significant differences in disease-free survival (DFS) or overall survival (OS) between the Girdin-negative and Girdin-positive breast cancer patients. a Kaplan-Meier estimates of DFS in nuclear Girdin (GN)-negative and GN-positive breast cancer patients. b Kaplan-Meier estimates of OS in GN-negative and GN-positive breast cancer patients. c Kaplan-Meier estimates of DFS in Girdin in the cytoplasm (GC)-negative and GC-positive breast cancer patients. d Kaplan-Meier estimates of OS in GC- negative and GC-positive breast cancer patients