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. 2021 Jan 20;12(1):102. doi: 10.1038/s41419-021-03393-5

Fig. 4. Identification of ANXA1 as a KIAA1199-binding membrane protein required for its HA-degrading activity.

Fig. 4

A Amino acid sequences of ANXA1 identified by LC-MS/MS (in blue rectangle). B The interaction of KIAA1199 with ANXA1 was analyzed by Co-IP using ANXA1 and KIAA1199 antibodies with the whole-cell lysates of RA FLS. C Colocalization of ANXA1 and KIAA1199 on RA FLS cell membrane by immunofluorescent staining. Images were captured with a confocal microscopy at a magnification of ×400. D Immunohistochemical staining of ANXA1 was performed on the sections of joint tissues from RA patients and normal subjects with the magnification of ×40 and ×100. E Western blot analysis of ANXA1 protein in FLS from normal subjects (n = 3) and RA patients (n = 3). F The effect of ANXA1 knockdown on fl-HA degradation mediated by membrane-bound sKIAA1199. G–I The effect of AXNA1 knockdown on the secretion of TNF-α (G), IL-1β (H), and IL-6 (I) by FLS from RA patients (n = 6). All reactions were conducted in triplicate and data were presented with mean ± SD.