<i>miR-200c</i> delivered using PEI nanoparticles inhibits IL-6, IL-8, and CCL-5 in primary human periodontal ligament fibroblasts.

<p><b>A-C</b>: The transcripts of IL-6 (<b>A</b>), IL-8 (<b>B</b>), and CCL-5 (<b>C</b>) in the cells with <i>miR-200c</i> or empty vector cultured in DMEM supplemented with LPS after 24 hours; <b>D</b> and <b>E</b>: the amounts of IL-6 (<b>D</b>), IL-8 (<b>E</b>), and CCL-5 (<b>F</b>) secreted by the cells with miR-200c or empty vector cultured in DMEM supplemented with LPS after 12 and 32 hrs, respectively. *: p<0.05 vs empty vector with the same amount.</p

Intracellular delivery of <i>miR-200c</i> using PEI nanoparticles to human primary periodontal ligament fibroblasts and bone marrow MSCs.

<p><b>A</b>: TEM image of PEI-<i>miR-200c</i> nanoplexes. <b>B</b> and <b>C:</b> Fold change of the transcript of <i>miR-200c</i> in non-treated human periodontal ligament fibroblasts (<b>B</b>) and bone marrow MSCs (<b>C</b>) and the cells transfected with empty vector (EV) (10μg/per well) and <i>miR-200c</i> (1, 5, 10μg/per well).</p

<i>miR-200c</i> overexpression in HEPM cells and the effects on their proliferation.

<p><b>A</b>: Microphotographs of HEPM cells and the cells with <i>miR-200c</i> or scrambled <i>miRs</i> under phase-contrast. Bar = 10μm. <b>B</b>: Fold change of <i>miR-200c</i> expression in non-treated HEPM cells and the cells with <i>miR-200c</i> and scrambled <i>miRs</i>. <b>C</b>: The doubling time of non-treated HEPM cells and the cells in the context of <i>miR</i> infection. **: p<0.01.</p

<i>miR-200c</i> increases osteogenic biomarkers in human preosteoblasts.

<p><b>A</b> and <b>B</b>: the amounts of the transcript of OCN (<b>A</b>) and calcium content (<b>B</b>) in non-treated HEPM cells and the cells with <i>miR-200c</i> or scrambled <i>miRs</i> cultured in DMEM supplemented <i>β</i>-glycerophosphate and ascorbic acid after 1 and 2 weeks, respectively. *: p<0.05.</p

<i>miR-200c</i> modulates proinflammatory mediators in human preosteoblasts.

<p><b>A</b> and <b>B:</b> the transcripts of IL-6 <b>(A)</b> and IL-8 (<b>B</b>) in non-treated HEPM cells and the cells with <i>miR-200c</i> or scrambled <i>miRs</i> cultured in DMEM supplemented with LPS at 0, 1, 5 and 10 μg/mL after 24 hours; *:p<0.05 vs non-treated; <b>C:</b> the amounts of IL-8 secreted by HEPM cells with <i>miR-200c</i> or scrambled <i>miRs</i> cultured in DMEM supplemented with or without LPS at different time points; *: p<0.05 vs cells with scrambled miRs; <b>D</b> and <b>E:</b> the amounts of IL-6 (<b>D</b>) and CCL-5 (<b>E</b>) secreted by HEPM cells with <i>miR-200c</i> or <i>scrambled miRs</i> cultured in DMEM supplemented with or without LPS after 24 hrs; <b>F:</b> the amounts of OPG secreted by HEPM cells with different <i>miRs</i> cultured in DMEM supplemented with or without LPS after 32 hours. *: p<0.05.</p

Enhancement of osteogenic differentiation of human bone marrow MSCs with overexpression of <i>miR-200c</i> using PEI nanoparticles.

<p><b>A</b>: Images of ALP and von-Kossa staining in MSCs overexpressing <i>miR-200c</i>, one and two weeks after treatment with osteogenic medium. <b>B</b> and <b>C</b>: the transcripts of ALP (<b>B</b>) and Runx2 (<b>C</b>) in MSCs overexpressing <i>miR-200c</i>, one week after treatment with osteogenic medium. <b>D</b> and <b>E</b>: Quantitative measurement of ALP levels (<b>D</b>) and calcium content (<b>E</b>) in MSCs overexpressing <i>miR-200c</i>, one and two week after treatment with osteogenic medium. Each measurement was made in triplicate. *: p<0.05.</p

<i>PMIS-200c</i> reduces binding activity of <i>miR-200c</i> to the <i>3’UTR</i> of IL-6, IL-8, and CCL-5 and the function of <i>miR-200c</i>.

<p><b>A-C</b>: Normalized luciferase activities of the <i>3’ UTR</i> IL-6, IL-8, and CCL-5-luciferase reporters and their <i>3’UTR</i>-mutated-luciferase reporters co-treated with <i>miR-200c</i> and <i>PMIS-EV</i> or PMIS-200c at different ratios of concentration. <b>D-F</b>: the transcripts of IL-6 (<b>D</b>), IL-8 (<b>E</b>), and CCL-5 (<b>F</b>) in the cells co-treated with <i>miR-200c</i> and <i>PMIS-EV</i> or <i>PMIS-200c</i> cultured in DMEM supplemented with LPS after 24 hours; *: p<0.05.</p

Additional file 6: of Genomics of NSCLC patients both affirm PD-L1 expression and predict their clinical responses to anti-PD-1 immunotherapy

Table S5. Analysis of the Discovery and Validation datasets was performed using Weka 3. The first number in each column represented the number of patient treatment responses correctly classified by the model. The second number represented the number of incorrectly classified patient treatment responses. The GOAL row at the bottom of each column described the number of correctly and incorrectly classified patients in the simulation models. The Test Set columns described the output from applying the model trained on the Discovery set to the Validation set. The “Test and Train” columns described test set accuracy (test set column) plus the training error (results obtained by applying the model to the training set, i.e. training error). (DOCX 19 kb

Additional file 8: of Genomics of NSCLC patients both affirm PD-L1 expression and predict their clinical responses to anti-PD-1 immunotherapy

Table S6. Comparisons of clinical and predicated responses and match scores. We used a cross-validation approach to assess the match scores in Table 1 of the PD-1 predicted responses against the PD-1 clinical responses in the Rizvi et al. 2015 Discovery dataset vs. the Validation dataset. We then pooled and re-partitioned the dataset into two new Training and Test datasets. We then used a similar cross-validation approach to assess the match scores of the PD-1 predicted responses vs. the PD-1 clinical responses. (DOCX 17 kb

Additional file 2: of Genomics of NSCLC patients both affirm PD-L1 expression and predict their clinical responses to anti-PD-1 immunotherapy

Table S2. Individual mutational profiles of patient drug responders (n = 11) and nonresponders (n = 18). (DOCX 19 kb