Journal: Nucleic Acids Research

Article Title: MicroRNA-target pairs in human renal epithelial cells treated with transforming growth factor β1: a novel role of miR-382

doi: 10.1093/nar/gkq718

Figure Lengend Snippet: Proteins differentially expressed in HK2 cells in response to TGFβ1

Article Snippet: Superoxide dismutase 2 (SOD2) human cDNA ORF clone was obtained from OriGene.

Techniques: Membrane

Journal: Nucleic Acids Research

Article Title: MicroRNA-target pairs in human renal epithelial cells treated with transforming growth factor β1: a novel role of miR-382

doi: 10.1093/nar/gkq718

Figure Lengend Snippet: Verification of proteomic results. HK2 cells were treated as described in . Protein samples used for the western blot analysis were independent of those used for the proteomic analysis. TPM1, tropomyosin 1; SOD2, superoxide dismutase 2. The gel shown for TPM1 was a time course analysis. Only the 48-h time point was plotted in the bar graph for comparison with the proteomic data. n = 4 for proteomics, n = 3 for western blot; *significantly different from vehicle ( P < 0.05 for western blot; see ‘Materials and Methods’ section for the criteria of differential expression for the proteomic analysis).

Article Snippet: Superoxide dismutase 2 (SOD2) human cDNA ORF clone was obtained from OriGene.

Techniques: Western Blot, Comparison, Quantitative Proteomics

Journal: Nucleic Acids Research

Article Title: MicroRNA-target pairs in human renal epithelial cells treated with transforming growth factor β1: a novel role of miR-382

doi: 10.1093/nar/gkq718

Figure Lengend Snippet: MiRNA-target pairs supported by sequence characteristics and reciprocal expression in human renal epithelial cells treated with TGFβ1

Article Snippet: Superoxide dismutase 2 (SOD2) human cDNA ORF clone was obtained from OriGene.

Techniques: Sequencing, Expressing

Journal: Nucleic Acids Research

Article Title: MicroRNA-target pairs in human renal epithelial cells treated with transforming growth factor β1: a novel role of miR-382

doi: 10.1093/nar/gkq718

Figure Lengend Snippet: miR-382 targeted SOD2 and contributed to TGFβ1-induced loss of epithelial characteristics in human renal epithelial cells. ( A ) TGFβ1 induced loss of epithelial characteristics in HK2 cells as indicated by typical changes in cell morphology and a suppression of E-cadherin mRNA, an epithelial marker. TGFβ1 was used at 3 ng/ml for 48 h. N = 6, * P < 0.05. ( B ) Knockdown of miR-382. HK2 cells were treated with vehicle or TGFβ1 in the presence of LNA anti-miR-382 or LNA scrambled control anti-miR (100 nM). n = 6, * P < 0.05 vs. no treatment; # P < 0.05 versus TGFβ1 plus control anti-miR. ( C ) Knockdown of miR-382 attenuated the suppression of E-cadherin mRNA by TGFβ1. n = 6–9, * P < 0.05 versus control anti-miR. ( D) miR-382 interacted with the 3′-UTR of SOD2 in Hela cells. Hela cells were transfected with luciferase reporter constructs containing a 3′-UTR segment of SOD2 or a mutated (SOD2-mut) or partially deleted (SOD2-del) segment. The mutations and deletion were described in the text. The effect of anti-miR-382 (100 nM) compared to control, scramble anti-miR was shown. n = 4, * P < 0.05 versus control anti-miR. ( E) miR-382 interacted with 3′-UTR of SOD2 in HK2 cells. HK2 cells were transfected with luciferase reporter constructs as described above. The effect of pre-miR-382 (100 nM), a miR-382 mimic, compared with control, scramble pre-miR was shown. n = 4, * P < 0.05 versus control pre-miR. ( F ) SOD2 expression was suppressed by TGFβ1, but partially restored by knockdown of miR-382. miR-382 in TGFβ1-treated HK2 cells was knocked down with LNA anti-miR (see B). n = 6, * P < 0.05 vs. control anti-miR.

Article Snippet: Superoxide dismutase 2 (SOD2) human cDNA ORF clone was obtained from OriGene.

Techniques: Marker, Knockdown, Control, Transfection, Luciferase, Construct, Expressing

Journal: Nucleic Acids Research

Article Title: MicroRNA-target pairs in human renal epithelial cells treated with transforming growth factor β1: a novel role of miR-382

doi: 10.1093/nar/gkq718

Figure Lengend Snippet: 3′-UTR reporter constructs

Article Snippet: Superoxide dismutase 2 (SOD2) human cDNA ORF clone was obtained from OriGene.

Techniques:

Journal: Nucleic Acids Research

Article Title: MicroRNA-target pairs in human renal epithelial cells treated with transforming growth factor β1: a novel role of miR-382

doi: 10.1093/nar/gkq718

Figure Lengend Snippet: SOD2 was protective against TGFβ1-induced loss of epithelial characteristics. ( A ) Overexpression of SOD2 with a Myc-DDK-tagged plasmid (pSOD2). HK2 cells were not transfected (NT) or were transfected with pSOD2 or an empty vector (2 µg for each 3.5-cm dish). Western blot was performed 24 h later using an SOD2 antibody. ( B ) Overexpression of SOD2 attenuated TGFβ1-induced downregulation of E-cadherin. n = 5, * P < 0.05.

Article Snippet: Superoxide dismutase 2 (SOD2) human cDNA ORF clone was obtained from OriGene.

Techniques: Over Expression, Plasmid Preparation, Transfection, Western Blot

Journal: eBioMedicine

Article Title: Single-nucleus RNA sequencing reveals heterogenous microenvironments and specific drug response between cervical squamous cell carcinoma and adenocarcinoma

doi: 10.1016/j.ebiom.2023.104846

Figure Lengend Snippet: Characteristics and functional status of T cells and macrophages in CSCC and CAde. (a – b) t-SNE plots of the T cells, colored by cell type and individual samples, respectively. (c) Heatmap displayed the expression profiles of marker genes in each identified T cell subcluster. (d) Box plots displayed the composition comparison of each T cell subcluster in CSCC and CAde. The P -values were calculated by Wilcoxon Rank Sum Test. (e) Violin plots depicted the estimated scores of different functional statues in identified T cell subclusters. ∗ P < 0.05, ∗∗∗ P < 0.001. The P -values were calculated by Wilcoxon Rank Sum Test. (f) Box plots displayed the cytotoxicity (left) and the exhausted (right) score of CD8 + T cells in CSCC and CAde. ∗∗∗ P < 0.001. The P -values were calculated by Wilcoxon Rank Sum Test. (g) Violin plot indicated the cytotoxicity scores of CSCC and CAde from the TCGA cohort. ∗ P < 0.05. The P -value was calculated by Wilcoxon Rank Sum Test. (h) Kaplan–Meier curve of overall survival based on the gene signature of cytotoxicity in all CC cohort from the TCGA database. The P -value was calculated by Log-rank test. (i) t-SNE plots of the myeloid cells, colored by cell type. (j) The fraction of each macrophage subcluster in eight individual samples. (k) Dot plots depicted the enrichment GO terms of each identified macrophage subcluster. (l) Heatmap displayed the functional activity scores of each macrophage subcluster based on known gene signatures. (m) Violin plot depicted the phagocytosis scores of CSCC and CAde. ∗∗∗ P < 0.001. The P -value was calculated by Wilcoxon Rank Sum Test. (n) mIF staining of macrophages with phagocytotic function in collected CSCC (n = 32) and CAde (n = 11) samples. (o) Box plots displayed the number of Macro-C2 (MRC1 + /CD68 + ) cells/field in CSCC and CAde. ∗∗∗ P < 0.001. The P -value was calculated by Wilcoxon Rank Sum Test.

Article Snippet: The details of primary antibodies used in this study were as follows: MTSS1 (Atlas Antibodies, #HPA075540, 1:200), POLA1 (Atlas Antibodies, #HPA002947, 1:200), POSTN (Abcam, #ab215199, 1:500), COL1A1 (CST, #72026, 1:100), LMCD1 (Immunoway, #YT7409, 1:100), MRC1 (CST, #91992, 1:100), CD68 (MXB, #kit-0026, 1:1), PROX1 (Proteintech, #67834-1-Ig, 1:250), SOD2 (Proteintech, #CL594-66474, 1:100) ( ).

Techniques: Functional Assay, Expressing, Marker, Comparison, Activity Assay, Staining