Alteration of chondroitin sulfate composition on proteoglycan produced by knock-in mouse embryonic fibroblasts whose versican lacks the A subdomain

Figures & data

Figure 1.  Agarose gel electrophoresis of PCR products from mouse embryo tails. A: wild-type allele determination by genomic PCR; B: mutant allele determination by genomic PCR. The numbers in figures A and B represent embryo as follows: number 1 = embryo no. 1; number 2 = embryo no. 2; number 3 = embryo no. 3; number 4 = embryo no. 4; number 5 = embryo no. 5; number 6 = embryo no. 6; number 7 = embryo no. 7; number 8 = embryo no. 8; number 9 = embryo no. 9; number 10 = embryo no. 10; M = 1 kb DNA ladder; number 11 = embryo no. 11; number 12 = embryo no. 12; number 13 = embryo no. 13; number 14 = embryo no. 14; number 15 = embryo no. 15; number 16 = embryo no. 16; number 17 = embryo no. 17; number 18 = embryo no. 18; number 19 = embryo no. 19; number 20 = embryo no. 20; number 21 = embryo no. 21; number 22 = embryo no. 22.

Figure 2.  Representative photographs (4× magnification) showing the cell morphology of wild-type (WT) fibroblasts (A) and Cspg2^Δ3/Δ3 fibroblasts (B) at passage no. 4.

Figure 3.  Versican deposition in WT and Cspg2^Δ3/Δ3 fibroblast cultures. The deposition of versican molecules was detected by the [³⁵S] incorporation assay; the amount of versican deposition was represented as the intensity of [³⁵S] isotope (cpm×10⁶). The closed bar (CL) represents the amount of versican deposited in cell layer, and the open bar (M) represents the amount of versican deposited in the culture medium. Data are the mean value±standard deviation of three independent experiments. WT = wild-type fibroblasts; Cspg2^Δ3/Δ3=Cspg2^Δ3/Δ3 (mutant) fibroblasts.

Table I.  Chondroitin sulfate composition in cell layer and fibroblast culture medium; data shown are the percent content of chondroitin sulfate. Data are the mean values of three independent experiments±standard deviation.

	% Chondroitin sulfate content
	Cell layer		Medium
	Wild type	Cspg2	Wild type	Cspg2
Δdi0S	13±0.11	19±0.79	33±0.50	28±0.30
Δdi4S	87±0.11	81±0.11	59±0.55	59±0.22
Δdi6S	ND	ND	8±0.05	12±0.33
ΔdiSE	ND	ND	ND	1±0.09
ΔdiSD	ND	ND	ND	ND
ΔdiTriS	ND	ND	ND	ND

Δdi0S: non-sulfate chondroitin sulfate, Δdi4S: chondroitin 4-sulfate, Δdi6S: chondroitin 6-sulfate, ΔdiSE: chondroitin 4,6-sulfate, ΔdiSD:chondroitin 2,6-sulfate, ΔdiTriS: chondroitin 2,4,6-sulfate.

Figure 4.  Chondroitin sulfate content on versican core protein which deposited in cell layer (upper graph) and culture medium (lower graph) of fibroblasts using high performance liquid chromatography (HPLC) analysis. Data are the mean values of three independent experiments±standard deviation. Each bar represents the chondroitin sulfate isomer.