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Fig. S1 PIPKI is highly expressed in cancer cell lines
Actin B2B H358 H441 PC9 H1975 A549 H1299 H460 SPC-A1 95D B PIPKI Actin siNC siI-1 siI-2 siI-3 C
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Fig. S2 Smurf1 degrades PIPKI
B Flag-Smurf1 PIPKIi2 PIPKIi1 GFP HA-PIPKI Flag-Smurf1 Flag-Smurf1 HA-PIPKIi2 HA-PIPKIi2 KD GFP Flag-Smurf1
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Fig. S3 Smurf1 interacts with PIPKI
B HA-PIPKIi Flag-Smurf1 HA-PIPKI IP: HA WCE HA-PIPKIi HA-PIPKIa HA-PIPKIb GFP-Ii2 WT GFP-PIPKIi2 IP: GFP WCE F-Smurf1 Talin GFP-Ii2 S650D GFP-Ii2 S650A C HA-PIPKIi F-Smurf1 WT HA-PIPKIi2 IP: HA WCE F-Smurf1 T306D
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Fig. S4 Lys-255 in PIPKIi2 is important for interacting with Smurf1
B HA-PIPKIi2 WT HA-PIPKIi2 K255R F-Smurf1CA HA-PIPKIi2 IP: HA WCE HA-PIPKIi2 WT HA-PIPKIi2 K255R F-talin Head Flag-talin head HA-PIPKIi2 IP: HA WCE
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Fig. S5 Smurf1 phosphorylation by PKA prevents Smurf1-mediated PIPKIg degradation
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Supplementary table 1: PIPKI is highly expressed in adenocarcinoma and squamous carcinoma.
Table1. Analysis of the association between PIPKI expression in membrane and clinicopathological features in NSCLC PIPKI Number Positive p-value Sex Male 77 28（36.3%） Female 46 14(30.4%) 0.502 Tabacco use Non-smoking 60 20(33.3%) Smoking 63 22(35%) 0.293 Pathology Adenocarcinoma 75 21(28%) Squamous carcinoma 48 19(39.6%) 0.151 Tumor size ≥3cm 70 27(38.6%) <3cm 53 14(26.4%) 0.178 N-status N0 13(18.6%) N1-2 28(52.8%) <0.0001 Stage Ⅰ 51 9(17.6%) Ⅱ 24 6(25%) Ⅲ 43 24(55.8%) Ⅳ 5 3(60%) 0.001 Histologic differentiation Poor 24(50%) Well/Moderate 17(22.7%) 0.002 Lympho-vascular invasion Absent 93 27(29.0%) Present 30 14(46.7%) < Pleural invasion 38 14(36.8%) 85 27(31.8%) 0.611
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Mutations Identified, n (%)
Table 2 Distribution and Frequency of Mutations in adenocarcinoma Mutations Identified, n (%) ASC (n=72) 95% CI (%) EGFR exon 19 deletion 20(27.8%) EGFR L858R 21(29.2%) 19-41 EGFR L861Q 2(2.8%) other mutationsa EGFR wild type 27(37.5%) aIncluded two double EGFR mutation, one was G719C,S768I and the other was S768I,G719A Table3 The association of PIPKI expression and EGFR mutation status in adenocarcinoma of lung Total(n=72) EGFR Mut. p-value PIPKI(+) 51 36(70.6%) PIPKI(-) 21 9(42.8%) 0.027
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