Journal: Molecular Medicine Reports

Article Title: MicroRNA-21-5p protects melanocytes via targeting STAT3 and modulating Treg/Teff balance to alleviate vitiligo

doi: 10.3892/mmr.2020.11689

Figure Lengend Snippet: Imbalance of T effector/Treg cells and dysregulation of miR-21-5p in patients with VIT. Percentages of (A) Th1, (B) Th17, (C) Th22 and (D) Treg cells in CD4 + T cells of peripheral blood mononuclear cells in patients with VIT and healthy controls (n=15 for each group) were measured via flow cytometry. Protein levels of (E) IFN-γ, (F) IL-17A and (G) IL-22 in the peripheral blood of patients with VIT and healthy controls were measured via enzyme-linked immunosorbent assay. Relative expression levels of (H) Foxp3 and (I) miR-21-5p in the peripheral blood of patients with VIT and healthy controls were measured via reverse transcription-quantitative PCR. GAPDH and U6 were used as the internal controls, respectively. **P<0.01; ***P<0.001. VIT, vitiligo; Th, T helper; IFN, interferon; IL, interleukin; miR, microRNA; Treg cells, regulatory T cells.

Article Snippet: CD4 + T cells used in the experiments were purified using CD4 microbeads (cat. no. 130-045-101; Miltenyi Biotec, Inc.) for PBMCs in peripheral blood with 95% purity.

Techniques: Flow Cytometry, Enzyme-linked Immunosorbent Assay, Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Molecular Medicine Reports

Article Title: MicroRNA-21-5p protects melanocytes via targeting STAT3 and modulating Treg/Teff balance to alleviate vitiligo

doi: 10.3892/mmr.2020.11689

Figure Lengend Snippet: miR-21-5p mimic inhibits STAT3 expression in T helper 17-polarized CD4 + T cells of patients with vitiligo. (A) Relative expression levels of (A) miR-21-5p and (B) STAT3 of the control, MC+NC, M+NC, MC+STAT3 and M+STAT3 groups were measured via reverse transcription-quantitative PCR. U6 and GAPDH were used as internal controls, respectively. *P<0.05 and ***P<0.001 vs. MC+NC; ^^ P<0.01 and ^^^ P<0.001 vs. MC+STAT3; ### P<0.001 vs. M+NC. (C) Relative STAT3 protein expression of the control, MC+NC, M+NC, MC+STAT3 and M+STAT3 groups were measured via western blot analysis. GAPDH was used as the internal control. *P<0.05 vs. MC+NC; ***P<0.001 vs. MC+NC; ### P<0.001 vs. M+NC; ^^ P<0.01 vs. MC+STAT3. miR, microRNA; M, miR-21-5p mimic; MC, mimic control; NC, negative control.

Article Snippet: CD4 + T cells used in the experiments were purified using CD4 microbeads (cat. no. 130-045-101; Miltenyi Biotec, Inc.) for PBMCs in peripheral blood with 95% purity.

Techniques: Expressing, Control, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Negative Control

Journal: Molecular Medicine Reports

Article Title: MicroRNA-21-5p protects melanocytes via targeting STAT3 and modulating Treg/Teff balance to alleviate vitiligo

doi: 10.3892/mmr.2020.11689

Figure Lengend Snippet: miR-21-5p overexpression regulates T effector/Treg cell balance via STAT3. Percentages of (A) Th17, (B) Th22 and (C) Treg cells in CD4 + T cells of the control, MC+NC, M+NC, MC+STAT3 and M+STAT3 groups were measured via flow cytometry. Protein levels of (D) IL-17A and (E) IL-22 of T cells in the control, MC+NC, M+NC, MC+STAT3 and M+STAT3 groups were measured via enzyme-linked immunosorbent assay. (F) Relative Foxp3 expression in the control, MC+NC, M+NC, MC+STAT3 and M+STAT3 groups were measured via reverse transcription-quantitative PCR. GAPDH was used as the internal control. *P<0.05, **P<0.01 and ***P<0.001 vs. MC+NC; ^ P<0.05 and ^^ P<0.01 vs. MC+STAT3; ## P<0.01 and ### P<0.001 vs. M+NC. Th, T helper; IL, interleukin; miR, microRNA; Treg cells, regulatory T cells; M, miR-21-5p mimic; MC, mimic control; NC, negative control.

Article Snippet: CD4 + T cells used in the experiments were purified using CD4 microbeads (cat. no. 130-045-101; Miltenyi Biotec, Inc.) for PBMCs in peripheral blood with 95% purity.

Techniques: Over Expression, Control, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Negative Control

Journal: Molecular Medicine Reports

Article Title: MicroRNA-21-5p protects melanocytes via targeting STAT3 and modulating Treg/Teff balance to alleviate vitiligo

doi: 10.3892/mmr.2020.11689

Figure Lengend Snippet: miR-21-5p overexpression may protect melanocytes via regulating T effector/regulatory T cell balance through STAT3. (A) Apoptosis rates of melanocytes in the control, MC+NC, M+NC, MC+STAT3 and M+STAT3 groups were measured via flow cytometry. Protein levels of (B) IL-17A and (C) IL-22 in the lower chamber of Transwell inserts after co-culture of melanocytes and Th17-polarized CD4 + T cells in the control, MC+NC, M+NC, MC+STAT3 and M+STAT3 groups was measured via enzyme-linked immunosorbent assay. (D) Tyrosinase activity and (E) relative melanin content in melanocytes in the control, MC+NC, M+NC, MC+STAT3 and M+STAT3 groups were measured using a microplate reader. *P<0.05, **P<0.01 and ***P<0.001 vs. MC+NC; ^ P<0.05 and ^^ P<0.01 vs. MC+STAT3; ## P<0.01 and ### P<0.001 vs. M+NC. Th, T helper; IL, interleukin; miR, microRNA; M, miR-21-5p mimic; MC, mimic control; NC, negative control.

Article Snippet: CD4 + T cells used in the experiments were purified using CD4 microbeads (cat. no. 130-045-101; Miltenyi Biotec, Inc.) for PBMCs in peripheral blood with 95% purity.

Techniques: Over Expression, Control, Flow Cytometry, Co-Culture Assay, Enzyme-linked Immunosorbent Assay, Activity Assay, Negative Control

Journal: Molecular Medicine Reports

Article Title: MicroRNA-21-5p protects melanocytes via targeting STAT3 and modulating Treg/Teff balance to alleviate vitiligo

doi: 10.3892/mmr.2020.11689

Figure Lengend Snippet: Potential mechanism of the interaction between miR-21-5p and STAT3 in vitiligo. Low miR-21-5p expression may induce the differentiation of CD4 + T cells towards Teff cells, causing a pro-inflammatory response and thereby increasing apoptosis in melanocytes. However, miR-21-5p mimic treatment promoted the balance of Treg/Teff cells and thereby had a benefit on the survival of melanocytes. Th, T helper; IFN, interferon; IL, interleukin; miR, microRNA; Treg/Teff cells, regulatory T/effector T cells.

Article Snippet: CD4 + T cells used in the experiments were purified using CD4 microbeads (cat. no. 130-045-101; Miltenyi Biotec, Inc.) for PBMCs in peripheral blood with 95% purity.

Techniques: Expressing