Table 1. GRASP KO alters HS and CS synthesis enzymes at both the mRNA and protein levels.

WT and GRASP KO cells were analyzed by proteomic and RNA-seq analyses. Shown are the fold changes of the indicated GAG synthesis enzymes in GRASP KO cells compared to that in WT cells. Enzymes in bold were further tested by Western blot.

Enzymes	Proteomics				RNA-seq
Common Enzymes	55KO/WT	p-value	65KO/WT	p-value	log2 (55KO/WT)	p-value	log2 (65KO/WT)	p-value	log2 (DKO/WT)	p-value
XylT1	ND		ND		−0.009	NA	6.511E-04	NA	−0.007	NA
XylT2	ND		ND		0.100	0.770	−0.250	0.290	−0.100	0.740
GalT1 (B4GALT7)	ND		ND		0.210	0.440	−0.170	0.500	0.140	0.590
GalT2	ND		ND		−0.850	0.020	−0.410	0.340	−0.110	0.840
GlcAT1 (B3GAT3)	ND		ND		−0.370	0.210	−0.110	0.750	0.020	0.960
PAPSS 1	0.047	0.411	−0.093	0.082	3.733E-03	0.990	−0.060	0.790	−0.120	0.460
PAPSS 2	−0.825	1.854E-05	−0.873	2.289E-06	−0.910	7.920E-10	−0.690	2.269E-06	−0.690	3.599E-06
PAPST 1 (SLC35B2)	−0.141	0.034	−0.069	0.256	−0.120	0.750	0.380	0.080	0.400	0.060
PAPST 2 (SLC35B3)	ND		ND		0.290	0.190	0.070	0.800	0.370	0.050
Enzymes in HS synthesis
EXTL2	ND		ND		−0.170	0.420	0.320	0.030	0.090	0.670
EXTL3	ND		ND		0.330	0.120	0.130	0.620	0.290	0.160
EXTL1	ND		ND		−0.010	NA	−0.010	NA	−0.010	NA
EXT1	ND		ND		0.200	0.420	0.660	3.090E-05	0.180	0.430
EXT2	ND		ND		−0.080	0.790	−0.100	0.690	−0.230	0.240
NDST 1	ND		ND		0.230	0.260	0.120	0.600	−0.170	0.400
NDST 2	ND		ND		0.280	0.620	−0.030	0.960	−0.050	0.940
NDST 3	ND		ND		−0.210	0.700	−0.460	0.200	0.100	0.850
NDST 4	ND		ND		ND		ND		ND
HS C5-EP (HSGLCE)	ND		ND		ND		ND		ND
H2ST	ND		ND		ND		ND		ND
H3ST 1	ND		ND		ND		ND		ND
H3ST 2	ND		ND		ND		ND		ND
H3ST 3A	ND		ND		ND		ND		ND
H3ST 3B	ND		ND		ND		ND		ND
H3ST 4	ND		ND		ND		ND		ND
H3ST 5	ND		ND		ND		ND		ND
H3ST 6	ND		ND		ND		ND		ND
H6ST 1	ND		ND		ND		ND		ND
H6ST 2	ND		ND		ND		ND		ND
H6ST 3	ND		ND		ND		ND		ND
Enzymes in CS synthesis
GalNAcT1 (GALNT1)	−0.404	8.206E-05	−0.521	1.34308E-05	−0.521	2.856E-09	−0.884	3.668E-26	−1.135	7.254E-42
GalNAcT2 (GALNT2)	−0.095	0.094	0.096	0.111	−0.140	0.480	0.290	0.040	−0.290	0.040
CHPF2	0.159	0.368	0.197	0.227	0.040	0.950	−0.050	0.910	0.200	0.540
CHSY1	ND		ND		−0.880	8.023E-07	−0.693	9.417E-05	−0.579	1.442E-03
CHSY2	ND		ND		NA		NA		NA
CHSY3	ND		ND		−0.110	0.790	0.050	0.900	0.110	0.740
C4ST 1 (CHST11)	ND		ND		−0.184	0.472	0.163	0.461	−0.177	0.424
C4ST 2 (CHST12)	ND		ND		−0.092	0.732	−0.022	0.935	−0.026	0.925
C4ST 3 (CHST13)	ND		ND		−0.270	0.635	0.480	0.240	0.903	0.008
GalNAc4S-6ST (CHST15)	ND		ND		−0.083	0.866	−0.734	0.003	−0.678	0.007
C6ST 1 (CHST3)	ND		ND		−0.242	0.471	−0.641	0.005	−0.839	1.600E-04
C6ST 2 (CHST4)	ND		ND		0	NA	0	NA	0.013	NA
UST	ND		ND		ND		ND		ND
DS-epi 1 (DSE)	ND		ND		−0.715	6.146E-10	−1.274	3.265E-28	−0.788	7.330E-12
DS-epi 2 (DSEL)	ND		ND		0.721	8.811E-05	0.505	8.939E-03	−0.001	9.967E-01
D4ST (CHST14)	ND		ND		ND		ND		ND
Enzymes in HA synthesis
HAS1	ND		ND		−0.009	NA	−0.009	NA	−0.007	NA
HAS2	ND		ND		0.131	0.850	0.088	0.888	0.390	0.356
HAS3	ND		ND		0.009	0.992	−0.132	0.819	0.066	0.915

NA, data not available; ND, not detected.