Fig. 4: Manganese supplementation rescue GAG elongation in TMEM165-deficient cells. | Cell Death & Disease

Fig. 4: Manganese supplementation rescue GAG elongation in TMEM165-deficient cells.

From: TMEM165 a new player in proteoglycan synthesis: loss of TMEM165 impairs elongation of chondroitin- and heparan-sulfate glycosaminoglycan chains of proteoglycans and triggers early chondrocyte differentiation and hypertrophy

Fig. 4

A Detection of decorin in conditioned medium of wild-type and tmem165-knockout mouse ATDC5 cells transfected with the expression vector for decorin. (B) Detection of HA-tagged syndecan 4 in cell lysates of wild-type and tmem165-knockout mouse ATDC5 cells transfected with HA-tagged syndecan 4 expression vector and grown in the presence or absence of MnCl2 (1 µM). β-actin was used as loading control (n = 3). C SDS-PAGE and autoradiography of [35S]-sulfate radiolabelled GAG chains primed with 4MU-Xyl in wild-type and tmem165-mutant mouse ATDC5 cells grown in the presence or absence of MnCl2 (1 µM) (n = 3). One representative blot of three independent experiments is shown. D Immunofluorescence analysis of cell surface HSGAG chains using anti-HS specific antibodies (green) in wild-type and tmem165-knockout mouse ATDC5 cells cultured in the absence and presence of Mn2+, Ba2+, Ca2+, Co2+, Mg2+ and Zn2+. The nucleus was stained with DAPI (blue). Digital images were captured with an inverted microscope, Leica DM13000. Representative images from three independent experiments are shown. Scale bar: 50 μm.

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