Fig 1: The protein patterns and expression levels of LRG, S100A8, C9, FN, AGP1, and A1AT in crude plasma, MARS-14 flow-though (FT) and MARS-14 elute (EL) fractions of the pooled samples of healthy controls (HC), patients with non-metastatic CRC (NM) and patients with metastatic CRC (M). (A) Stain-free imaging of the gel displayed the pattern of total proteins from three groups (HC, NM, and M). Crude plasma (30 µg), flow-through (5 µg) and elute (5 µg) fractions of three groups were separated on 10% TGX stain-free FastCast. Total proteins were visualized by a stain-free imaging system. (B) Immunoblots of LRG, S100A8, C9, FN, AGP1, and A1AT. Values below immunoblots denote the ratio of each protein band intensity normalized by its total protein loading and compared to those of the healthy control.
Fig 2: Scatter plots show relative expression levels of (A) LRG, (B) S100A8, (C) C9, (D) FN, (E) AGP1, and (F) A1AT from immunoblotting of individual plasma samples in the study-cohort and validation-cohort. All immunoblot results are provided in the supplementary data, Table S4 and S5. Green dots, Healthy control (HC); Red dots, Non-Metastatic CRC patients (NM); Blue dots, Metastatic CRC patients (M); Black dots, all CRC patients (CRC). Black lines represent the medians of samples in each group. Stars represent statistical significance calculated by non-parametric one-way ANOVA (Kruskal-Wallis) and Dunn’s multiple comparison test for the comparison among HC, NM and M groups; and non-parametric t-test (Mann-Whitney U test) for the comparison between HC and CRC groups. *, **, and *** represent p-value < 0.05, < 0.01, and < 0.001, respectively.
Fig 3: ORM1 was essential to cell proliferation. (a) The expression of ORM1 protein in A498, 786-O, and Caki-2 cells was much higher than the 293 T cells. (b) The gray value analysis of protein in (a). (c) ORM1 was knocked down in 786-O and Caki-2 cells. (d) The gray value analysis of protein in (c). (e) Cell proliferation of 786-O cells was inhibited in ORM1-KD group compared to NC group. (f) Cell proliferation of Caki-2 cells was inhibited in ORM1-KD group compared to NC group. #p < 0.05 showed statistically difference.
Fig 4: ORM1 enhanced the efficiency of sorafenib in KIRC. (a) Sorafenib inhibited cell proliferation in concentration-dependent manner. (b) The efficiency of sorafenib was enhanced in ORM1-KD group but relieved in ORM1-OE group in 786-O cells. (c) The efficiency of sorafenib was enhanced in ORM1-KD group but relieved in ORM1-OE group in Caki-2 cells. #p < 0.05 showed statistically difference.
Fig 5: ORM1 regulated cell apoptosis through CALR in KIRC. (a) Cell apoptosis was detected by Annexin V/FITC dye method. (b) The statistical analysis of cells in cell migration in (a). #p < 0.05 showed statistically difference.
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