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. 2022 Oct 11;18(7):1591–1600. doi: 10.4103/1673-5374.355979

Figure 5.

Figure 5

circ_0002538 regulates the expression of PLLP in vitro and in vivo.

(A) We examined the mRNA expression of the myelination-related genes SERINC5, PLLP, GJC3, PLP1, PRX, and MPZ in the circ_0002538-overexpressing SCs. Y-axis: fold changes in mRNA expression compared with the vector group. *P < 0.05, **P < 0.01, ***P < 0.001, vs. vector group (independent-sample t-test). (B) We used western blot analysis to evaluate the effect of circ_0002538 on PLLP in SCs. (C, D) We examined PLLP expression in the SCs cultured in ox-LDL via RT-PCR and western blotting. Y-axis: fold changes in PLLP mRNA expression compared with the 0 μg/mL group. *P < 0.05, ***P < 0.001, vs. 0 μg/mL group (one-way analysis of variance and Tukey’s post hoc test). (E, F) We evaluated the effect of circ_0002538 on PLLP in SCs cultured with ox-LDL via RT-PCR and western blotting. Y-axis: fold changes in PLLP mRNA expression compared with the control group. **P < 0.01, vs. control group; #P < 0.05, vs. SCs cultured in ox-LDL (one-way analysis of variance and Tukey’s post hoc test). (G, H) We tested PLLP protein expression in peripheral nerve tissues from patients with or without DPN via western blotting (n = 6). Y-axis: Fold changes in PLLP protein expression normalized to β-actin compared with the vector group. **P < 0.01 (independent-sample t-test). (I) The overexpression of circ_0002538 increased the protein expression level of PLLP in the sciatic nerve of mice with DPN (n = 3). (J) Quantification of PLLP by the densitometry of protein bands. *P < 0.05, vs. vector group (paired t-test). All bar graphs represent the average of three independent replicates, and the data are given as the mean ± SD. DPN: Diabetic peripheral neuropathy; GJC3: gap junction protein gamma 3; MPZ: myelin protein zero; ox-LDL-C: oxidized low-density lipoprotein cholesterol; PLLP: plasmolipin; PLP1: proteolipid protein 1; PRX: periaxin; SERINC5: serine incorporator 5; sh-circ_0002538: short hairpin RNA for circ_0002538; sh-NC: normal control for short hairpin RNA.fffc