Journal: BMC Plant Biology

Article Title: Identification of microRNAs in developing wheat grain that are potentially involved in regulating grain characteristics and the response to nitrogen levels

doi: 10.1186/s12870-020-2296-7

Figure Lengend Snippet: Length distribution of miRNAs in developing wheat grain of plants grown under two nitrogen application levels and sampled at 7, 17, and 27 DAA. HN-7, HN-17 and HN-27 represent grain at 7, 17, and 27 DAA under high nitrogen application levels, respectively. LN-7, LN-17 and LN-27 represent grain at 7, 17, and 27 DAA under low nitrogen application levels, respectively

Article Snippet: A SYBR PrimeScript miRNA RT-PCR Kit was used to perform qRT-PCR in a fluorescence detection system (TianGen Biotech, Beijing, China) following the manufacturer’s instructions.

Techniques:

Journal: BMC Plant Biology

Article Title: Identification of microRNAs in developing wheat grain that are potentially involved in regulating grain characteristics and the response to nitrogen levels

doi: 10.1186/s12870-020-2296-7

Figure Lengend Snippet: Venn diagram of differentially expressed miRNAs in different libraries at three grain developmental stages under high (HN) and low (LN) nitrogen treatment. HN-7, HN-17 and HN-27 represent grain at 7, 17, and 27 days after anthesis under high nitrogen application levels, respectively. LN-7, LN-17 and LN-27 represent grain at 7, 17, and 27 days after anthesis under low nitrogen application levels, respectively. The number outside (inside) the brackets represents the number of total (novel) differentially expressed miRNAs

Article Snippet: A SYBR PrimeScript miRNA RT-PCR Kit was used to perform qRT-PCR in a fluorescence detection system (TianGen Biotech, Beijing, China) following the manufacturer’s instructions.

Techniques:

Journal: BMC Plant Biology

Article Title: Identification of microRNAs in developing wheat grain that are potentially involved in regulating grain characteristics and the response to nitrogen levels

doi: 10.1186/s12870-020-2296-7

Figure Lengend Snippet: The major GO categories of “cellular component”, “biological process”, and “molecular function” for the predicted genes of all the differentially expressed miRNAs during grain development

Article Snippet: A SYBR PrimeScript miRNA RT-PCR Kit was used to perform qRT-PCR in a fluorescence detection system (TianGen Biotech, Beijing, China) following the manufacturer’s instructions.

Techniques:

Journal: BMC Plant Biology

Article Title: Identification of microRNAs in developing wheat grain that are potentially involved in regulating grain characteristics and the response to nitrogen levels

doi: 10.1186/s12870-020-2296-7

Figure Lengend Snippet: Conserved miRNAs differentially expressed in developing wheat grain and their predicted target gene functions

Article Snippet: A SYBR PrimeScript miRNA RT-PCR Kit was used to perform qRT-PCR in a fluorescence detection system (TianGen Biotech, Beijing, China) following the manufacturer’s instructions.

Techniques: Ubiquitin Proteomics, Activity Assay, Protein Binding, Binding Assay, Alternative Splicing

Journal: BMC Plant Biology

Article Title: Identification of microRNAs in developing wheat grain that are potentially involved in regulating grain characteristics and the response to nitrogen levels

doi: 10.1186/s12870-020-2296-7

Figure Lengend Snippet: Differentially expressed novel miRNAs associated with grain development and their target gene functions

Article Snippet: A SYBR PrimeScript miRNA RT-PCR Kit was used to perform qRT-PCR in a fluorescence detection system (TianGen Biotech, Beijing, China) following the manufacturer’s instructions.

Techniques: Cell Differentiation, Ubiquitin Proteomics, Protein Binding

Journal: BMC Plant Biology

Article Title: Identification of microRNAs in developing wheat grain that are potentially involved in regulating grain characteristics and the response to nitrogen levels

doi: 10.1186/s12870-020-2296-7

Figure Lengend Snippet: Relationships within a miRNA-gene-GO network for cell differentiation, seed development and response to heat stress as determined by Cytoscape. The red, green, and yellow colors indicate miRNAs, target gene functions and GO terms, respectively

Article Snippet: A SYBR PrimeScript miRNA RT-PCR Kit was used to perform qRT-PCR in a fluorescence detection system (TianGen Biotech, Beijing, China) following the manufacturer’s instructions.

Techniques: Cell Differentiation

Journal: BMC Plant Biology

Article Title: Identification of microRNAs in developing wheat grain that are potentially involved in regulating grain characteristics and the response to nitrogen levels

doi: 10.1186/s12870-020-2296-7

Figure Lengend Snippet: Differentially expressed miRNAs between HN and LN treatments and their predicted target gene functions

Article Snippet: A SYBR PrimeScript miRNA RT-PCR Kit was used to perform qRT-PCR in a fluorescence detection system (TianGen Biotech, Beijing, China) following the manufacturer’s instructions.

Techniques: Ubiquitin Proteomics, Alternative Splicing, Activity Assay

Journal: BMC Plant Biology

Article Title: Identification of microRNAs in developing wheat grain that are potentially involved in regulating grain characteristics and the response to nitrogen levels

doi: 10.1186/s12870-020-2296-7

Figure Lengend Snippet: Model for miRNAs and their target genes associated with grain development and the response to N levels. The black letters in green circles indicate miRNAs, and the red letters in black boxes indicate the target gene function. The black arrows represent the direction of regulation. TF, transcription factor; CRF, cytokinin response factor; SCF, Skp-cullin-F-box; GLU, glucosyltransferase; BRI, brassinosteroid insensitive; ASR, alternative splicing regulator; LRR-RPK, leucine-rich-repeat receptor-like protein kinase; K1P-PSS1, kinesin-1-like protein PSS1; ARF, auxin response factor; HNT, high-affinity nitrate transport; Redox, reduction-oxidation; HSP, heat-shock protein; POD, peroxidase

Article Snippet: A SYBR PrimeScript miRNA RT-PCR Kit was used to perform qRT-PCR in a fluorescence detection system (TianGen Biotech, Beijing, China) following the manufacturer’s instructions.

Techniques: Alternative Splicing

Journal: BMC Plant Biology

Article Title: Identification of microRNAs in developing wheat grain that are potentially involved in regulating grain characteristics and the response to nitrogen levels

doi: 10.1186/s12870-020-2296-7

Figure Lengend Snippet: Verification of the expression patterns of nine miRNAs present in developing wheat grain. The different lowercase letters above the columns indicate significant differences ( P < 0.05)

Article Snippet: A SYBR PrimeScript miRNA RT-PCR Kit was used to perform qRT-PCR in a fluorescence detection system (TianGen Biotech, Beijing, China) following the manufacturer’s instructions.

Techniques: Expressing

Journal: BMC Plant Biology

Article Title: Identification of microRNAs in developing wheat grain that are potentially involved in regulating grain characteristics and the response to nitrogen levels

doi: 10.1186/s12870-020-2296-7

Figure Lengend Snippet: Verification of the expression patterns of miRNA target genes in developing wheat grain. The different lowercase letters above the columns indicate significant differences ( P < 0.05)

Article Snippet: A SYBR PrimeScript miRNA RT-PCR Kit was used to perform qRT-PCR in a fluorescence detection system (TianGen Biotech, Beijing, China) following the manufacturer’s instructions.

Techniques: Expressing

Journal: Advanced Science

Article Title: Hippo Pathway Activation in Aged Mesenchymal Stem Cells Contributes to the Dysregulation of Hepatic Inflammation in Aged Mice

doi: 10.1002/advs.202300424

Figure Lengend Snippet: Downregulation of YAP1 in aged MSCs leads to reduced immunosuppressive function. A GSEA analysis of gene expression of young and aged MSCs shows that Hippo signaling is upregulated in aged MSCs. B) mRNA expression levels of Yap1 and YAP1 target genes were tested by RT‐PCR. C,D) Cytosolic and nuclear expression of YAP1 was detected by western blotting and quantified relative to the internal controls β‐ACTIN and Histone H3. E,F) Translocation of YAP1 into the nucleus was detected in young and aged MSCs by immunofluorescence staining and corresponding quantification. G) YAP1 expression was tested in young and aged liver MSCs by western blotting. β‐ACTIN was used as the internal control. H,I) IHC staining and quantification of CD8 + T‐cells in the liver. J,K) H&E staining of the liver after treatment with young and aged MSCs, with or without manipulation of Yap1 expression, and quantification of the necrotic areas. L) Measurement of serum ALT and AST levels in the indicated mice. Y‐MSCs, young MSCs; A‐MSCs, aged MSCs. N.D., not detected. Data (B, D, F) were analyzed using two‐tailed unpaired Student t ‐test. Data (I, K, L) were analyzed using one‐way ANOVA test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Graphs showed mean ± S.D. N = 3, the experiments were repeated for three times.

Article Snippet: A Bestar qPCR RT Kit (DBI Bioscience, Ludwigshafen, Germany) was used for reverse transcription to cDNA, and real‐time PCR was performed using a SYBR PrimeScript RT‐PCR Kit (DBI bioscience, Ludwigshafen, Germany) with an ABI Prism 7300 system.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Translocation Assay, Immunofluorescence, Staining, Immunohistochemistry, Two Tailed Test

Journal: Advanced Science

Article Title: Hippo Pathway Activation in Aged Mesenchymal Stem Cells Contributes to the Dysregulation of Hepatic Inflammation in Aged Mice

doi: 10.1002/advs.202300424

Figure Lengend Snippet: YAP1 maintains the immunosuppressive activity of MSCs by promoting iNOS expression. A) mRNA expression levels of genes encoding anti‐inflammation factors were detected in young and aged MSCs by RT‐PCR after treatment with I+T for 0 h, 4 h, and 8 h. B) Expression of iNOS protein in young and aged MSCs was detected by western blotting assay at 0, 24, and 48 h after I+T treatment and corresponding quantification. β‐ACTIN was used as the internal control. C, Nitrate concentrations were measured by a Griess reagent kit in conditioned medium of young and aged MSCs after I+T treatment. D,E) IHC staining of CD8 positive cells in the liver after treatment with young and aged MSCs (with or without manipulation of Nos2 expression) and corresponding quantification. F,G) H&E staining of the liver after treatment with young and aged MSCs (with or without manipulation of No` expression) and quantification of the necrotic areas. H,I) Young MSCs were transfected by shYap1 adenovirus, and aged MSCs were transfected by Yap1 overexpression adenovirus. iNOS expression was detected by western blotting assay in young and aged MSCs treated with I+T for the indicated time. Y‐MSCs, young MSCs; A‐MSCs, aged MSCs. N.D., not detected. Data (A, B, C) were analyzed using two‐tailed unpaired Student t ‐test. Data (E, G) were analyzed using one‐way ANOVA test. *** p < 0.001, **** p < 0.0001. Graphs showed mean ± S.D. N = 3, the experiments were repeated for three times.

Article Snippet: A Bestar qPCR RT Kit (DBI Bioscience, Ludwigshafen, Germany) was used for reverse transcription to cDNA, and real‐time PCR was performed using a SYBR PrimeScript RT‐PCR Kit (DBI bioscience, Ludwigshafen, Germany) with an ABI Prism 7300 system.

Techniques: Activity Assay, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Immunohistochemistry, Staining, Transfection, Over Expression, Two Tailed Test

Journal: Advanced Science

Article Title: Hippo Pathway Activation in Aged Mesenchymal Stem Cells Contributes to the Dysregulation of Hepatic Inflammation in Aged Mice

doi: 10.1002/advs.202300424

Figure Lengend Snippet: YAP1 promotes iNOS expression by binding the Stat1 promoter and enhancing Stat1 transcription. A, B, Young MSCs were transfected by shYap1 adenovirus and then treated with I+T for 0, 30, and 60 min. Aged MSCs were transfected by Yap1 overexpression adenovirus and then treated with I+T for 0, 30, and 60 min. Expression of YAP1, STAT1, and p‐STAT1 (Ser727) was detected by western blotting. β‐ACTIN was used as the internal control. C,D) Detection of iNOS expression by western blotting assay in young and aged MSCs in the rescue experiment. β‐ACTIN was used as the internal control. E) The mRNA expression levels of Stat1 in young and aged MSCs were detected by RT‐PCR after adenovirus‐mediated manipulation of Yap1 expression. F) Young MSCs were treated with and without 10 µM VP for 24 h. The levels of STAT1, CTGF, and CYR61 were detected by western blotting assay. β‐ACTIN was used as the internal control. G) CUT&Tag assay was performed to verify the target motif bound by YAP1/TEADs in the Stat1 promoter. Anti‐IgG was used as the negative control. Y‐MSCs, young MSCs; A‐MSCs, aged MSCs. Data were analyzed using two‐tailed unpaired Student t ‐test. ** p < 0.01, *** p < 0.001. The graph showed mean ± S.D. N = 3, the experiments were repeated for three times.

Article Snippet: A Bestar qPCR RT Kit (DBI Bioscience, Ludwigshafen, Germany) was used for reverse transcription to cDNA, and real‐time PCR was performed using a SYBR PrimeScript RT‐PCR Kit (DBI bioscience, Ludwigshafen, Germany) with an ABI Prism 7300 system.

Techniques: Expressing, Binding Assay, Transfection, Over Expression, Western Blot, Reverse Transcription Polymerase Chain Reaction, Negative Control, Two Tailed Test