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. 2017 Jan 5;7:39869. doi: 10.1038/srep39869

Figure 3. Effects of SDS on RSC 96 via SEM, immunohistochemical analysis, Western Blot assay and gene expression analysis in vitro.

Figure 3

(A) Scanning electron microscopy (SEM) of the cells on scaffolds at 4 days. (PLGA means cultured with 0 mM SDS, PLGA+S-0.1 means cultured with 0.1 mM SDS, PLGA+S-0.2 means cultured with 0.2 mM SDS, PLGA+S-0.4 means cultured with 0.4 mM SDS, TCP means cultured on TCP alone, TCP+s-0.2 means cultured with 0.2 mM SDS on TCP). Statistic analysis of scanning electron microscopy (SEM). Different letters denote significances with P < 0.05 and the same letter shows no significant differences (P ≥ 0.05). (B) Immunohistochemical analysis for S-100 protein, RSC 96 showed positive cytoplasmic staining for S-100 at 4 days. (PLGA means cultured with 0 mM SDS, PLGA+S-0.1 means cultured with 0.1 mM SDS, PLGA+S-0.2 means cultured with 0.2 mM SDS, PLGA+S-0.4 means cultured with 0.4 mM SDS, TCP means cultured on TCP alone, TCP+s-0.2 means cultured with 0.2 mM SDS on TCP). Statistic analysis of immunohistochemical analysis. Different letters denote significances with P < 0.05 and the same letter shows no significant differences (P ≥ 0.05). (C) Western Blot assay of S-100 protein and quantification of the proteins expression. Full-length blots are presented in supplementary information. Different letters denote significances with P < 0.05 and the same letter shows no significant differences (P ≥ 0.05); (D) Gene expression analysis of important neurotrophic factors (GDNF, BDNF and CNTF) by qRT-PCR in six groups at 2, 4 and 6 days. The gene expression levels were analyzed by the 2−ΔΔCT method using GAPDH as the internal control. The data represent the mean of three independent experiments (n = 3, mean ± SEM). Different letters denote significances with P<0.05 and the same letter shows no significant differences (P ≥ 0.05). (PLGA means cultured with 0 mM SDS, PLGA+S-0.1 means cultured with 0.1 mM SDS, PLGA+S-0.2 means cultured with 0.2 mM SDS, PLGA+S-0.4 means cultured with 0.4 mM SDS, TCP means cultured on TCP alone, TCP+s-0.2 means cultured with 0.2 mM SDS on TCP).