Additional file 2: of Prevalence of type 2 diabetes, prediabetes, and gestational diabetes mellitus in women of childbearing age in Middle East and North Africa, 2000–2017: protocol for two systematic reviews and meta-analyses

Box S1. Databases and search terms. Table S2. Data extraction parameters. (DOCX 44 kb

Down-regulation of HDAC3 by Rg3 alters cell cycle protein expression.

<p>A375 cells were treated with HDAC3 siRNA, Rg3 (50 µg/mL), MS-275 (10 µM), or HDAC3 siRNA transfection followed by Rg3 treatment. Control: untransfected cells; Mock: cells transfected with vector. (A) Western blotting analysis of expression levels of PRB, cyclin E, cyclin D1, CDK4, CDK2 and p21. GAPDH was used as an internal control. (B) The statistical analyses for Western blotting results were shown. Data were presented as the mean±S.D. of three independent experiments. *, <i>p<</i>0.05; **, <i>p<</i>0.01.</p

Down-regulating HDAC3 expression inhibits melanoma cell proliferation.

<p>HDAC3 was detected by real-time PCR (A) and Western blotting (B) in A375 cells after tansfection with HDAC3 siRNAs. Cell viability was determined by CCK-8 assay in A375 and C8161 cells (C). PCNA protein expression in A375 and C8161 cells (D). Control: untransfected cells; Mock: cells transfected with vector; siHDAC3: cells transfected with HDAC3-1 or HDAC3-2 siRNA. Cells were exposed to MS-275 (5 µM, 10 µM) for 24 h. Cell viability was determined by CCK-8 assay (E). GAPDH was used as an internal control. The statistical analyses for Western blotting results were shown Data were presented as the mean±S.D. of three independent experiments. *, <i>P</i><0.05; **, <i>P</i><0.01.</p

Increased expression of HDAC3 correlates with lymph node metastasis and clinical stage of melanoma.

<p>Immunohistochemistry staining of HDAC3 in human melanoma tissues. Normal and cancer tissues were analyzed by H&E staining (a, b). Negative control of normal and cancer tissues (c, d). The expression of HDAC3 in normal and cancer tissues (e, f). Bar = 50 µm (magnification, ×200). Insets showed enlarged views (magnification, ×400).</p

Rg3 decreases HDAC3 expression.

<p>(A) Analysis of HDAC3 level by Western blotting in A375 and C8161 cells after cells treated with Rg3 (0, 25, 50 µg/mL) for 72 h. (B) HDAC3 expression level in cells treated with Rg3 (50 µg/mL) for 0, 24, 48 or 72 h. GAPDH was used as an internal control. The statistical analyses for Western blotting results were shown (*, <i>p<</i>0.05; <i>**, p<</i>0.01). (C) Immunofluorescent staining of HDAC3 in A375 cells after Rg3 (50 µg/mL) treatment for 0, 24, 48 or 72 h. DAPI used as a nuclear staining. Bar  = 20 µm (magnification, ×400). Data were presented as the mean±S.D. of three independent experiments.</p

Rg3 inhibits melanoma cell growth.

<p>(A) Structure of 20 (R)-Ginsenoside Rg3. (B) A375 and C8161 cells were treated with Rg3 (0, 25, 50, 100 µg/mL) for 24, 48 or 72 h. Cell viability was determined by CCK-8 assay as described in Materials and Methods. (C) Cell cycle analysis. A375 cells were treated with Rg3 (50 µg/mL) for 0, 24, 48 or 72 h and were analyzed by a FACScan flow cytometer. Group data analysis was shown. (D) The mRNA level of PCNA was detected by real-time PCR in A375 cells. (E) Western blotting was employed to examine PCNA protein expression in A375 cells. GAPDH was used as an internal control. The statistical analyses for Western blotting were shown. Data were presented as the mean±S.D. of three independent experiments. *, <i>p</i><0.05; **, <i>p</i><0.01.</p

Correlation of HDAC3 protein expression to clinicopathologic features of melanoma Patients.

<p>Abbreviations: CSD, melanomas on skin with chronic sun-induced damage;NSD, melanomas on skin without chronic sun-induced damage; UP, melanoma of unknown primary.</p><p>Correlation of HDAC3 protein expression to clinicopathologic features of melanoma Patients.</p

Rg3 and down-regulating HDAC3 expression increase p53 acetylation and transcription activity.

<p>Cells were treated with Rg3 (0, 50 µg/mL) for 24 h. (A) Western blotting analysis of the expression levels of Ac-p53 (k373/382) in A375, C8161 and SK-MEL-28 cells. (B) Immunofluorescent staining analysis of expression levels of Ac-p53 (k373/382) in A375 cells. Cells were transfected with HDAC3 siRNA (48 h) or exposed to MS-275 (10 µM) for 24 h. (C) Western blotting analysis the expression levels of Ac-p53 (k373/382) in A375, C8161 and SK-MEL-28 cells. (D) Immunofluorescent staining analysis the expression levels of Ac-p53 (k373/382) in A375 cells. Control: untransfected cells; Mock: cells transfected with vector; siHDAC3: cells transfected with HDAC3 siRNA. Bar = 20 µm (magnification, ×400). GAPDH was used as an internal control. The statistical analyses for Western blotting results were shown. (E) A375 and C8161 cells were transfected with p53-luciferase reporter plasmid. After 24 h, the cells were treated with Rg3 (50 µg/mL) or MS-275 (10 µM) for 24 h, or transfection with siHDAC3 for 24 h. Luciferase activity was measured in cell lysates. Data were presented as the mean±S.D. of three independent experiments. *, <i>p</i><0.05; **, <i>p</i><0.01.</p

Characteristics of Female Migrants (n = 555) in Al Ain, United Arab Emirates, 2014.

<p>Characteristics of Female Migrants (n = 555) in Al Ain, United Arab Emirates, 2014.</p

Cases of T2DM, prevalence and odds ratios^* for selected independent variables.

<p>Cases of T2DM, prevalence and odds ratios^{<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0169949#t002fn001" target="_blank">*</a>} for selected independent variables.</p