Fig 1: Reciprocal regulation of AnxA6 and GRF2 by non-selective calcium channel blockers.A) BT-549 (AnxA6 high) and empty vector or AnxA6 expressing HCC1806 (AnxA6-low) cells were treated with Ni2+ or Ca2+ for 72 h and AnxA6, AnxA2 and GRF2 protein assessed by immunoblotting. B) Densitometric analysis of AnxA6 and GRF2 expression. Bars represent fold change relative to control cells from two independent experiments. C and D) Effects of Ni2+ concentration and time on the reciprocal expression of AnxA6 and GRF2. AnxA6 expressing HCC1806 cells were treated with the indicated concentrations of Ni2+ for 72 h (C) or at 0.5 mM Ni2+ for the indicated times (D); Densitometric analysis of the expression of AnxA6, GRF2 and AnxA2 from a representative experiment (C and D, left panels). E) HCC1806 cells were treated with the indicated concentrations of bepridil or amlodipine for 72 h and the expression of AnxA6 and GRF2 assessed by western blotting. F and G) Densitometric analysis of the protein bands in bepridil treated (F) and amlodipine treated cells from a representative experiment (G). Flag-AnxA6: flag-tagged recombinant AnxA6, End-AnxA6: endogenously expressed AnxA6.
Fig 2: Reciprocal expression of AnxA6 and GRF2 in TNBC PDX models.A) Whole tissue extracts were prepared from patient derived xenografts (n = 19), and equal amounts of protein were analyzed by western blotting using antibodies to the indicated proteins. B) Densitometric analysis of the expression of the indicated proteins. Shown are protein band intensities normalized to β-actin from a representative experiment. C) The ratio of GRF2:AnxA6 was used to classify the tumors into potentially rapidly growing or invasive tumors as in Fig 6. D) A plot of the reciprocal expression of AnxA6 and GRF2 based on the GRF2:AnxA6 ratio for each PDX sample and showing two major groups: Group 1 with a strong potential for rapid growth and Group 3 with a strong potential for invasiveness, separated by a more diverse Group 2 with varied potentials for growth and invasiveness.
Fig 3: Correlation between RASGRF2 transcript level and Immune infiltration pattern. (A) Varying predicted tumor infiltration proportions of 24 immune subtypes. (B-E) Correlations between RASGRF2 transcript level and predicted immune infiltration levels. (F-I) Comparison of immune infiltration levels between high and low RASGRF2 transcript level groups (P < 0.001).
Fig 4: The basic workflow of a comprehensive study is outlined. The study comprises two-part. (A) The transcript expression signature, prognostic and diagnostic value of RASGRF2. (B) The functional value, tumor immune infiltration analysis, clinical use, and preliminary validation of RASGRF2 in STAD.
Fig 5: Prognostic significance of RASGRF2 transcript level. (A) Kaplan-Meier survival analysis for patients with stomach adenocarcinoma exhibiting high versus low RASGRF2 transcript levels (P = 0.023). (B) A five-year nomogram based on prognostic risk factors was identified by multivariate Cox regression. (C) Nomogram calibration plots.
Supplier Page from Abcam for Anti-RASGRF2 antibody