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. 2016 Nov;11(11):1830–1838. doi: 10.4103/1673-5374.194755

Figure 5.

Figure 5

Knockdown of Rab27a and Slp2-a in Schwann cells reduces myelin protein expression and myelin-like membrane formation in the DRG neuron and Schwann cell co-cultures.

(A–D) Compared with the non-targeting control siRNA, Rab27a and Slp2-a knockdown in the purified Schwann cells prior to seeding with DRG neurons significantly reduces mRNA levels (A) and protein expression (C) of myelin proteins MAG, P0, and PMP22 at 14 days after co-culture. Optical density results are presented as a fold of gapdh expression (B) and actin expression (D). (E) Double-immunostaining of MBP and NF in different groups at 14 days in co-culture also shows that silencing of Rab27a and Slp2-a reduces the formation of MAG-positive myelin-like membranes. Scale bar: 50 μm. (F) Quantification of MBP immunofluorescence intensity in different groups at 14 days after co-culture. Student's t-test, n = 3 cultures/group. All data represent mean ± SEM. *P < 0.05 (one-way analysis of variance followed by the Newman-Keuls test). DRG: Dorsal root ganglion; MAG: myelin associated glycoprotein; NF: neurofilament; PMP22: peripheral myelin protein 22; MBP: myelin basic protein; P0: protein zero; GAPDH: glyceraldehyde-3-phosphate dehydrogenase.