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. 2022 Aug 2;18(3):634–642. doi: 10.4103/1673-5374.350210

Figure 5.

Figure 5

Apoptosis and demyelination in the spinal cord at L2/3 level caused by the absence of SPARC.

(A) TUNEL staining with NeuN and immunofluorescence staining of βIII-tubulin (green), Bax (red), and cleaved caspase-3 (red). TUNEL-positive cells were observed and apoptosis-related proteins Bax and cleaved caspase-3 expression were increased in SPARC-null mouse neurons compared with WT mice. Scale bar: 50 µm in TUNEL staining; 20 µm in others. (B) TUNEL staining with Olig2 (red) and immunofluorescence staining of CNPase (green) with cleaved caspase-3 (red). TUNEL-positive cells were observed in SPARC-null mice oligodendrocytes and cleaved caspase-3 expression was increased in SPARC-null mice oligodendrocytes compared with WT mice. Scale bar: 100 µm in TUNEL staining; 50 µm in other panels. (C) Immunofluorescence staining for MBP (green). MBP was less integrated in SPARC-null mice compared with WT mice. Scale bar: 50 µm. (D) Western blot and quantitative analysis (normalized to β-actin) of Bax, Bcl-2, and cleaved caspase-3. Data are shown as mean ± SD (n = 3 mice in each group). **P < 0.01, ****P < 0.0001 (Student’s t-test). CNPase: 2-3-Cyclic nucleotide 3-phosphodiesterase; MBP: myelin basic protein; NeuN: neuronal nuclei; Olig2: oligodendrocyte transcriptional factor-2; SPARC: secreted protein, acidic and rich in cysteine; Tunel: terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-nick end labeling staining; WT: wild-type.