Figure 1

CSPG inhibits the adhesion and migration of GRPs.

(a) RT-PCR analysis showing the expression of PTPRS in cultured GRPs using two specific primer pairs. RT⁺ indicates PCR with reverse transcription product as template. RT⁻ indicates the parallel negative control reaction without reverse transcriptase, as template. (b) Immunofluorescence staining of cultured GRPs with anti-PTPRS (red) and anti-GFAP (green) antibodies. Nuclei were counterstained with DAPI (blue). Note that both GFAP+ and GFAP- GRPs express PTPRS. Scale bar, 75 μm. (c) Quantification of the number of GRP cell aggregates on cover glass treated with different substrates. Con: PLL (poly-l-lysine, 100 μg/mL) coated cover glass; CSPG: CSPG (3 μg/mL) treatment after coating with PLL; Chase: Treatment with Chase conditioned medium after CSPG treatment. **p < 0.005 (t-test). (d–g) representative images showing the effect of different treatments on the migration of GRPs from the cell aggregate. GRPs were incubated in suspension for 1 day to allow the formation of cell aggregates, which were plated onto cover glass coated with CSPG or laminin (15 μg/mL). CSPG/Chase: cover glass was coated with CSPG, followed by treatment with Chase condition medium overnight before plating GRP cell aggregates; CSPG/laminin: Cover glass was coated with CSPG (3 μg/mL) and laminin (15 μg/mL) was applied into the culture medium. All cover glass were pretreated with PLL. (h) Cumulative distribution of the diameter of GRP aggregates after overnight culture on different substrates. (D) Dose effect of CSPG on GRP migration from cell aggregates and the mitigation of CSPG inhibition by bath application of laminin (10 μg/mL). Data are mean ± SEM.