Structured Review

TaKaRa ajehhadh
Effect of estivation on the relative expression levels of <t>AjEHHADH</t> . (A) Relative mRNA expression levels of AjEHHADH in the intestine of NA and DA groups respectively. Values were normalized against β -Tubulin. “*” indicates significant statistical differences ( P
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1) Product Images from "The potential contribution of miRNA-200-3p to the fatty acid metabolism by regulating AjEHHADH during aestivation in sea cucumber"

Article Title: The potential contribution of miRNA-200-3p to the fatty acid metabolism by regulating AjEHHADH during aestivation in sea cucumber

Journal: PeerJ

doi: 10.7717/peerj.5703

Effect of estivation on the relative expression levels of AjEHHADH . (A) Relative mRNA expression levels of AjEHHADH in the intestine of NA and DA groups respectively. Values were normalized against β -Tubulin. “*” indicates significant statistical differences ( P
Figure Legend Snippet: Effect of estivation on the relative expression levels of AjEHHADH . (A) Relative mRNA expression levels of AjEHHADH in the intestine of NA and DA groups respectively. Values were normalized against β -Tubulin. “*” indicates significant statistical differences ( P

Techniques Used: Expressing

The complete cDNA sequence and deduced amino acid sequence of AjEHHADH from A. japonicus. Amino acids are numbered starting with the N-terminal Met residue. The asterisk indicates the translational termination codon. The open reading frame (ORF) from the initiation codon (ATG) to the termination codon (TAG) is notated by uppercase letters. At the bottom of the page is the schematic diagram of domains and characteristic motifs.
Figure Legend Snippet: The complete cDNA sequence and deduced amino acid sequence of AjEHHADH from A. japonicus. Amino acids are numbered starting with the N-terminal Met residue. The asterisk indicates the translational termination codon. The open reading frame (ORF) from the initiation codon (ATG) to the termination codon (TAG) is notated by uppercase letters. At the bottom of the page is the schematic diagram of domains and characteristic motifs.

Techniques Used: Sequencing, Polyacrylamide Gel Electrophoresis

Functional analysis of miR-200-3p and AjEHHADH in vivo . (A) Relative AjEHHADH mRNA transcripts expression after transfection with miRNA modified mimics. (B) Western blot analysis after transfection with miRNA modified mimics. “*” indicates significant differences ( P
Figure Legend Snippet: Functional analysis of miR-200-3p and AjEHHADH in vivo . (A) Relative AjEHHADH mRNA transcripts expression after transfection with miRNA modified mimics. (B) Western blot analysis after transfection with miRNA modified mimics. “*” indicates significant differences ( P

Techniques Used: Functional Assay, In Vivo, Expressing, Transfection, Modification, Western Blot

Identification and characterization of the miRNA-200-3p binding sites in the 3′ UTR of AjEHHADH and functional effect of miR-200-3p on AjEHHADH . (A) Schematic representation of the putative miRNA-200-3p targeting sites in AjEHHADH mRNA and the respective mutant sites. (B) HEK-293T cells were co-transfected with the pMIRREPORT-BHMT-WT vector, carrying the wild-type and the mutated AjEHHADH 3′-UTR, pRLCMV-Renilla-luciferase, and control miR-200-3p mimics as indicated. “*”indicates significant differences ( P
Figure Legend Snippet: Identification and characterization of the miRNA-200-3p binding sites in the 3′ UTR of AjEHHADH and functional effect of miR-200-3p on AjEHHADH . (A) Schematic representation of the putative miRNA-200-3p targeting sites in AjEHHADH mRNA and the respective mutant sites. (B) HEK-293T cells were co-transfected with the pMIRREPORT-BHMT-WT vector, carrying the wild-type and the mutated AjEHHADH 3′-UTR, pRLCMV-Renilla-luciferase, and control miR-200-3p mimics as indicated. “*”indicates significant differences ( P

Techniques Used: Binding Assay, Functional Assay, Mutagenesis, Transfection, Plasmid Preparation, Luciferase

Theoretical binding of miR-200-3p to a conserved region in the 3′ UTR of the AjEHHADH gene. (A) Conservation analysis of the miR-200-3p binding site in the EHHADH gene from sea cucumber A. japonicus and Lingula anatina . The seed region sequence (shaded) shows 100% conservation between these two sequences. (B) Predicted binding structure of miR-200-3p when binding to the 3′ UTR of AjEHHADH, as determined from the TargetScan and miRanda program.
Figure Legend Snippet: Theoretical binding of miR-200-3p to a conserved region in the 3′ UTR of the AjEHHADH gene. (A) Conservation analysis of the miR-200-3p binding site in the EHHADH gene from sea cucumber A. japonicus and Lingula anatina . The seed region sequence (shaded) shows 100% conservation between these two sequences. (B) Predicted binding structure of miR-200-3p when binding to the 3′ UTR of AjEHHADH, as determined from the TargetScan and miRanda program.

Techniques Used: Binding Assay, Sequencing

2) Product Images from "Combined small RNA and gene expression analysis revealed roles of miRNAs in maize response to rice black-streaked dwarf virus infection"

Article Title: Combined small RNA and gene expression analysis revealed roles of miRNAs in maize response to rice black-streaked dwarf virus infection

Journal: Scientific Reports

doi: 10.1038/s41598-018-31919-z

Different expressed known miRNAs identified in sRNA libraries.
Figure Legend Snippet: Different expressed known miRNAs identified in sRNA libraries.

Techniques Used:

Function classification of the target genes of novel miRNAs.
Figure Legend Snippet: Function classification of the target genes of novel miRNAs.

Techniques Used:

Potential regulatory roles of miRNAs and their targets in maize response to RBSDV.
Figure Legend Snippet: Potential regulatory roles of miRNAs and their targets in maize response to RBSDV.

Techniques Used:

Number of different expressed miRNAs in response to RBSDV.
Figure Legend Snippet: Number of different expressed miRNAs in response to RBSDV.

Techniques Used:

qRT-PCR verification of miRNAs and genes.
Figure Legend Snippet: qRT-PCR verification of miRNAs and genes.

Techniques Used: Quantitative RT-PCR

3) Product Images from "CircPSMC3 suppresses the proliferation and metastasis of gastric cancer by acting as a competitive endogenous RNA through sponging miR-296-5p"

Article Title: CircPSMC3 suppresses the proliferation and metastasis of gastric cancer by acting as a competitive endogenous RNA through sponging miR-296-5p

Journal: Molecular Cancer

doi: 10.1186/s12943-019-0958-6

CircPSMC3 expression is down regulated in clinical GC specimens and cell lines. a Scatter plot analyses of circRNAs microarray data showing differentially expressed circRNA in lymph node metastasis group and normal group . High expression level is indicated by “red” and low levels by “green”. b Scatter plot analyses of circRNA microarray data showing differentially expressed circRNAs in lymph node metastasis group and no lymph node metastasis group. c GO analysis of circRNAs in lymph node metastasis group and normal group. d GO analysis of circRNAs in lymph node metastasis group and no lymph node metastasis group. e The spliced mature sequence length of circPSMC3 derived from the PSMC3 gene is 502 bp. f QRT-PCR for the abundance of circPSMC3 in GC cells treated with RNase R. g QRT-PCR for the abundance of PSMC3 mRNA in GC cells treated with RNase R. h CircPSMC3 expressions were evaluated using qRT-PCR in 106 pairs of gastric cancer plasmas and 21 pairs of normal plasmas. i CircPSMC3 expressions were measured using qRT-PCR in 106 pairs of gastric cancer (Tumor) and matched noncancerous tissue (Normal). j The expressions of circPSMC3 were evaluated in cell lines using qRT-PCR. k The area under the ROC curve (AUC) in distinguishing GC plasmas and normal ones was 0.9326. l Kaplan Meier survival curve showed the relationship between circPSMC3 and survival rates. ** p
Figure Legend Snippet: CircPSMC3 expression is down regulated in clinical GC specimens and cell lines. a Scatter plot analyses of circRNAs microarray data showing differentially expressed circRNA in lymph node metastasis group and normal group . High expression level is indicated by “red” and low levels by “green”. b Scatter plot analyses of circRNA microarray data showing differentially expressed circRNAs in lymph node metastasis group and no lymph node metastasis group. c GO analysis of circRNAs in lymph node metastasis group and normal group. d GO analysis of circRNAs in lymph node metastasis group and no lymph node metastasis group. e The spliced mature sequence length of circPSMC3 derived from the PSMC3 gene is 502 bp. f QRT-PCR for the abundance of circPSMC3 in GC cells treated with RNase R. g QRT-PCR for the abundance of PSMC3 mRNA in GC cells treated with RNase R. h CircPSMC3 expressions were evaluated using qRT-PCR in 106 pairs of gastric cancer plasmas and 21 pairs of normal plasmas. i CircPSMC3 expressions were measured using qRT-PCR in 106 pairs of gastric cancer (Tumor) and matched noncancerous tissue (Normal). j The expressions of circPSMC3 were evaluated in cell lines using qRT-PCR. k The area under the ROC curve (AUC) in distinguishing GC plasmas and normal ones was 0.9326. l Kaplan Meier survival curve showed the relationship between circPSMC3 and survival rates. ** p

Techniques Used: Expressing, Microarray, Sequencing, Derivative Assay, Quantitative RT-PCR

4) Product Images from "A sucrose non‐fermenting‐1‐related protein kinase‐1 gene, IbSnRK1, improves starch content, composition, granule size, degree of crystallinity and gelatinization in transgenic sweet potato"

Article Title: A sucrose non‐fermenting‐1‐related protein kinase‐1 gene, IbSnRK1, improves starch content, composition, granule size, degree of crystallinity and gelatinization in transgenic sweet potato

Journal: Plant Biotechnology Journal

doi: 10.1111/pbi.12944

Diagram showing the regulation of starch biosynthesis in the storage roots of the IbSnRK1‐overexpressing sweet potato plants. Biosynthesis pathways are shown with solid arrows and regulatory interactions are shown with broken arrows. Fold changes (the mean of the four transgenic lines L13, L14, L17 and L18) are shown in colour, red boxes, white boxes and blue boxes indicate up‐regulation, no obvious change and down‐regulation of expression of genes encoding these enzymes (proteins), respectively.
Figure Legend Snippet: Diagram showing the regulation of starch biosynthesis in the storage roots of the IbSnRK1‐overexpressing sweet potato plants. Biosynthesis pathways are shown with solid arrows and regulatory interactions are shown with broken arrows. Fold changes (the mean of the four transgenic lines L13, L14, L17 and L18) are shown in colour, red boxes, white boxes and blue boxes indicate up‐regulation, no obvious change and down‐regulation of expression of genes encoding these enzymes (proteins), respectively.

Techniques Used: Transgenic Assay, Expressing

Expression analysis of IbSnRK1 (a), starch content (b) and amylose proportion (c) in the storage roots of the transgenic sweet potato plants, WT and VC. The sweet potato β‐actin gene was used as an internal control. Data are presented as the mean ± SD ( n = 3). * and ** indicate a significant difference compared with WT at P
Figure Legend Snippet: Expression analysis of IbSnRK1 (a), starch content (b) and amylose proportion (c) in the storage roots of the transgenic sweet potato plants, WT and VC. The sweet potato β‐actin gene was used as an internal control. Data are presented as the mean ± SD ( n = 3). * and ** indicate a significant difference compared with WT at P

Techniques Used: Expressing, Transgenic Assay

Expression analysis of IbSnRK1 in sweet potato. (a) Starch content and IbSnRK1 expression in the storage roots of different cultivars. (b) Expression of IbSnRK1 in different tissues of Lushu 3. L, leaf; P, petiole; S, stem; SR, storage root; FR, fibrous root. (c) Expression of IbSnRK1 in response to 175 m m sucrose treatment. The treatment with H 2 O was used as control. (d) Time settings used to examine the circadian rhythm during 16‐h light/8‐h dark photoperiods with light on at 5 am and off at 9 pm. Total RNA was extracted from the in vitro grown plants sampled at 10 pm (P1, P4, P7 and P10), 10 am (P2, P5, P8 and P11) and 4 pm (P3, P6, P9 and P12), respectively. The results are expressed as relative values with respect to P1 (set to 1.0). Data are presented as the mean ± SD ( n = 3). * and different lowercase letters indicate a significant difference at P
Figure Legend Snippet: Expression analysis of IbSnRK1 in sweet potato. (a) Starch content and IbSnRK1 expression in the storage roots of different cultivars. (b) Expression of IbSnRK1 in different tissues of Lushu 3. L, leaf; P, petiole; S, stem; SR, storage root; FR, fibrous root. (c) Expression of IbSnRK1 in response to 175 m m sucrose treatment. The treatment with H 2 O was used as control. (d) Time settings used to examine the circadian rhythm during 16‐h light/8‐h dark photoperiods with light on at 5 am and off at 9 pm. Total RNA was extracted from the in vitro grown plants sampled at 10 pm (P1, P4, P7 and P10), 10 am (P2, P5, P8 and P11) and 4 pm (P3, P6, P9 and P12), respectively. The results are expressed as relative values with respect to P1 (set to 1.0). Data are presented as the mean ± SD ( n = 3). * and different lowercase letters indicate a significant difference at P

Techniques Used: Expressing, In Vitro

5) Product Images from "Cytokinesis-required Cdc14 is a signaling hub of asexual development and multi-stress tolerance in Beauveria bassiana"

Article Title: Cytokinesis-required Cdc14 is a signaling hub of asexual development and multi-stress tolerance in Beauveria bassiana

Journal: Scientific Reports

doi: 10.1038/srep03086

Multi-phenotypic defects observed in Δ cdc14 versus two control strains. (A) Colony sizes after 6-day growth on minimal CZA (3% sucrose as mere carbon and 0.3% NaNO 3 as mere nitrogen) and CZA-derived media [altered carbon: glucose (Glu), galactose (Gal), glycerol (Gly), acetate (Ace) or carbon starvation (CS); altered nitrogen: NaNO 2 , NH 4 Cl or nitrogen starvation (NS)]. (B −D) Effective concentrations (EC 50 s) of stressful chemicals required for the suppression of 50% colony growth after 6-day incubation at 25°C on the plates of 1/4 SDAY supplemented with the gradients of H 2 O 2 , menadione (MND), NaCl, carbendazim (CBD), Congo red (CR) and SDS respectively. (E) Residue viability of conidia after 24 h incubation at 25°C on germination medium supplemented with NaCl (1.2 M), MND (0.2 mM), H 2 O 2 (4 mM), CR (0.5 mg/ml) or SDS (0.04%). (F) Median lethal time (LT 50 ) for conidial tolerance to wet-heat stress at 45°C and median lethal dose (LD 50 ) for conidial UV-B resistance. (G) LT 50 (no. days) for conidial virulence to S. litura second-instar larvae under a uniform spray. The asterisked bar in each group differs significantly from those unmarked (Tukey's HSD, P
Figure Legend Snippet: Multi-phenotypic defects observed in Δ cdc14 versus two control strains. (A) Colony sizes after 6-day growth on minimal CZA (3% sucrose as mere carbon and 0.3% NaNO 3 as mere nitrogen) and CZA-derived media [altered carbon: glucose (Glu), galactose (Gal), glycerol (Gly), acetate (Ace) or carbon starvation (CS); altered nitrogen: NaNO 2 , NH 4 Cl or nitrogen starvation (NS)]. (B −D) Effective concentrations (EC 50 s) of stressful chemicals required for the suppression of 50% colony growth after 6-day incubation at 25°C on the plates of 1/4 SDAY supplemented with the gradients of H 2 O 2 , menadione (MND), NaCl, carbendazim (CBD), Congo red (CR) and SDS respectively. (E) Residue viability of conidia after 24 h incubation at 25°C on germination medium supplemented with NaCl (1.2 M), MND (0.2 mM), H 2 O 2 (4 mM), CR (0.5 mg/ml) or SDS (0.04%). (F) Median lethal time (LT 50 ) for conidial tolerance to wet-heat stress at 45°C and median lethal dose (LD 50 ) for conidial UV-B resistance. (G) LT 50 (no. days) for conidial virulence to S. litura second-instar larvae under a uniform spray. The asterisked bar in each group differs significantly from those unmarked (Tukey's HSD, P

Techniques Used: Derivative Assay, Incubation

Disruption of cdc14 altered the transcripts of stress-responsive genes and the phosphorylation of Hog1 and Slt2. (A−D) Relative transcript levels (RTL) of stress-responsive genes in the 1/4 SDAY cultures of Δ cdc14 versus wild type supplemented with MND (menadione 0.2 mM), CR (Congo red 0.5 mg/ml), NaCl (0.8 M) and CBD (carbendazim 0.5 μg/ml) for 3-day growth at 25°C respectively (assessed via qRT-PCR with paired primers in Table S1 ). (E and F) OD 450 values for the respective phosphorylation levels of Hog1 and Slt2 in the protein extracts of fungal cultures grown under the same stresses (BC: unstressed blank control). The extracts were probed with phospho-p38 and phospho-p42/44 antibodies. The asterisked bar in each group differs significantly from those unmarked (Tukey's HSD, P
Figure Legend Snippet: Disruption of cdc14 altered the transcripts of stress-responsive genes and the phosphorylation of Hog1 and Slt2. (A−D) Relative transcript levels (RTL) of stress-responsive genes in the 1/4 SDAY cultures of Δ cdc14 versus wild type supplemented with MND (menadione 0.2 mM), CR (Congo red 0.5 mg/ml), NaCl (0.8 M) and CBD (carbendazim 0.5 μg/ml) for 3-day growth at 25°C respectively (assessed via qRT-PCR with paired primers in Table S1 ). (E and F) OD 450 values for the respective phosphorylation levels of Hog1 and Slt2 in the protein extracts of fungal cultures grown under the same stresses (BC: unstressed blank control). The extracts were probed with phospho-p38 and phospho-p42/44 antibodies. The asterisked bar in each group differs significantly from those unmarked (Tukey's HSD, P

Techniques Used: Quantitative RT-PCR

6) Product Images from "The Molecular Basis of Inactivation of Metronidazole-Resistant Helicobacter pylori Using Polyethyleneimine Functionalized Zinc Oxide Nanoparticles"

Article Title: The Molecular Basis of Inactivation of Metronidazole-Resistant Helicobacter pylori Using Polyethyleneimine Functionalized Zinc Oxide Nanoparticles

Journal: PLoS ONE

doi: 10.1371/journal.pone.0070776

Morphological transition and rRNA degradation. H. pylori incubated without or with ZnO-PEI NP for 3 h was visualized by (a) SEM and (b) TEM. (c) RNA was extracted from ZnO-PEI NP treated and untreated H. pylori and electrophoresed on 7M urea-6% polyacrylamide gel. In (a), (b) and (c) the left panel shows the control and the right panel shows treated cells. (d) 16S and 23S rRNA was estimated in the RNA samples by qRT-PCR (list of primers are provided in Table S2 ). * P
Figure Legend Snippet: Morphological transition and rRNA degradation. H. pylori incubated without or with ZnO-PEI NP for 3 h was visualized by (a) SEM and (b) TEM. (c) RNA was extracted from ZnO-PEI NP treated and untreated H. pylori and electrophoresed on 7M urea-6% polyacrylamide gel. In (a), (b) and (c) the left panel shows the control and the right panel shows treated cells. (d) 16S and 23S rRNA was estimated in the RNA samples by qRT-PCR (list of primers are provided in Table S2 ). * P

Techniques Used: Incubation, Transmission Electron Microscopy, Quantitative RT-PCR

7) Product Images from "Identification and Expression Profile Analysis of Odorant Binding Proteins in the Oriental Fruit Fly Bactrocera dorsalis"

Article Title: Identification and Expression Profile Analysis of Odorant Binding Proteins in the Oriental Fruit Fly Bactrocera dorsalis

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms140714936

Tissue distribution of the ten identified OBPs . qRT-PCR to analyze OBP transcript levels in different tissues of mature adults during the oviposition period, including head, thorax, abdomen, leg, wing, male antennae, and female antennae. Relative transcript levels are calculated using β -actin as the standard. Different letters indicate significant differences in the expression level of OBPs ( p
Figure Legend Snippet: Tissue distribution of the ten identified OBPs . qRT-PCR to analyze OBP transcript levels in different tissues of mature adults during the oviposition period, including head, thorax, abdomen, leg, wing, male antennae, and female antennae. Relative transcript levels are calculated using β -actin as the standard. Different letters indicate significant differences in the expression level of OBPs ( p

Techniques Used: Quantitative RT-PCR, Expressing

8) Product Images from "Naturally occurring antisense RNA of histone H2a in mouse cultured cell lines"

Article Title: Naturally occurring antisense RNA of histone H2a in mouse cultured cell lines

Journal: BMC Genetics

doi: 10.1186/1471-2156-6-23

(a) RT-PCR products from cDNAs obtained by priming total RNA with random hexamers. Lanes: 1, DNA ladder (100-bp ladder, TOYOBO); 2–7, RT-PCR products amplified with primers F1 and R1 (upper) and those amplified with primers F1 and R2 (lower). RNA was extracted from Hepa 1–6 (lanes 2 and 5), 3T3 (lanes 3 and 6), and LLC (lanes 4 and 7) cells. Superscript III was not added in the reaction of lanes 2–4. Lane 8, PCR product of genomic DNA amplified with primers F1 and R1 (upper) and that amplified with primers F1 and R2 (lower). Arrows indicate the expected products. (b) Patterns of digestion of PCR products by Pst I. Lanes: 1, DNA ladder; 2–4, digests of PCR products amplified with primers F1 and R1. PCR product was produced from Hepa 1–6 (lane 2), 3T3 (lane 3), and LLC (lane 4). Lanes 5–7, digests of PCR products amplified with primers F1 and R2. PCR product was produced from Hepa 1–6 (lane 5), 3T3 (lane 6), and LLC (lane 7). Arrows indicate the expected products. (c) RT-PCR products and the Eco RI-digest patterns of cDNAs obtained by priming total RNA with the specific primer R3. Lanes: 1 and 8, DNA ladder; 2–7, RT-PCR products amplified with primers F2 and R3. RNA was extracted from Hepa 1–6 (lanes 2 and 5), 3T3 (lanes 3 and 6), and LLC (lanes 4 and 7). Superscript III was not added in the reaction of lanes 2–4. Lanes 9–11, Eco RI-digests of PCR products amplified with primers F2 and R3. PCR product was produced from Hepa 1–6 (lane 9), 3T3 (lane 10), and LLC (lane 11). Arrows indicate the expected products.
Figure Legend Snippet: (a) RT-PCR products from cDNAs obtained by priming total RNA with random hexamers. Lanes: 1, DNA ladder (100-bp ladder, TOYOBO); 2–7, RT-PCR products amplified with primers F1 and R1 (upper) and those amplified with primers F1 and R2 (lower). RNA was extracted from Hepa 1–6 (lanes 2 and 5), 3T3 (lanes 3 and 6), and LLC (lanes 4 and 7) cells. Superscript III was not added in the reaction of lanes 2–4. Lane 8, PCR product of genomic DNA amplified with primers F1 and R1 (upper) and that amplified with primers F1 and R2 (lower). Arrows indicate the expected products. (b) Patterns of digestion of PCR products by Pst I. Lanes: 1, DNA ladder; 2–4, digests of PCR products amplified with primers F1 and R1. PCR product was produced from Hepa 1–6 (lane 2), 3T3 (lane 3), and LLC (lane 4). Lanes 5–7, digests of PCR products amplified with primers F1 and R2. PCR product was produced from Hepa 1–6 (lane 5), 3T3 (lane 6), and LLC (lane 7). Arrows indicate the expected products. (c) RT-PCR products and the Eco RI-digest patterns of cDNAs obtained by priming total RNA with the specific primer R3. Lanes: 1 and 8, DNA ladder; 2–7, RT-PCR products amplified with primers F2 and R3. RNA was extracted from Hepa 1–6 (lanes 2 and 5), 3T3 (lanes 3 and 6), and LLC (lanes 4 and 7). Superscript III was not added in the reaction of lanes 2–4. Lanes 9–11, Eco RI-digests of PCR products amplified with primers F2 and R3. PCR product was produced from Hepa 1–6 (lane 9), 3T3 (lane 10), and LLC (lane 11). Arrows indicate the expected products.

Techniques Used: Reverse Transcription Polymerase Chain Reaction, Amplification, Polymerase Chain Reaction, Produced

9) Product Images from "Induction of Epithelial Mesenchimal Transition and Vasculogenesis in the Lenses of Dbl Oncogene Transgenic Mice"

Article Title: Induction of Epithelial Mesenchimal Transition and Vasculogenesis in the Lenses of Dbl Oncogene Transgenic Mice

Journal: PLoS ONE

doi: 10.1371/journal.pone.0007058

qRT-PCR analysis of genes selected from the microarray expression profile. qRT-PCR analysis of genes selected from the microarray expression profile. For the 2, 14, and 42 day old Dbl transgenic and wild type mice lenses 2 µg of total RNA was reverse-transcribed and Real Time quantitative PCR (qRT-PCR) was conducted in triplicate for each target transcript. Expression changes of 24 selected genes were evaluated in relation to the values obtained in parallel for three reference genes. The results are expressed as log2 ratios of fold changes (Dbl relative to wild type lenses) and the mean of triplicate determinations for each target transcript is shown. Positive values indicate that the mRNA level of a particular gene is up-regulated, whereas negative values indicate that the transcript is down-regulated. The bars represent SE.
Figure Legend Snippet: qRT-PCR analysis of genes selected from the microarray expression profile. qRT-PCR analysis of genes selected from the microarray expression profile. For the 2, 14, and 42 day old Dbl transgenic and wild type mice lenses 2 µg of total RNA was reverse-transcribed and Real Time quantitative PCR (qRT-PCR) was conducted in triplicate for each target transcript. Expression changes of 24 selected genes were evaluated in relation to the values obtained in parallel for three reference genes. The results are expressed as log2 ratios of fold changes (Dbl relative to wild type lenses) and the mean of triplicate determinations for each target transcript is shown. Positive values indicate that the mRNA level of a particular gene is up-regulated, whereas negative values indicate that the transcript is down-regulated. The bars represent SE.

Techniques Used: Quantitative RT-PCR, Microarray, Expressing, Transgenic Assay, Mouse Assay, Real-time Polymerase Chain Reaction

10) Product Images from "The diagnostic value of circulating microRNAs for middle-aged (40–60-year-old) coronary artery disease patients"

Article Title: The diagnostic value of circulating microRNAs for middle-aged (40–60-year-old) coronary artery disease patients

Journal: Clinics

doi: 10.6061/clinics/2015(04)07

Plasma expression levels of miR-149, miR-424 and miR-765 in non-CAD patients, stable CAD patients and unstable CAD patients.
Figure Legend Snippet: Plasma expression levels of miR-149, miR-424 and miR-765 in non-CAD patients, stable CAD patients and unstable CAD patients.

Techniques Used: Expressing

Diagnostic accuracy of circulating miR-149, miR-424 and miR-765 were analyzed by ROC curve. A) ROC curve of miR-149, non-CAD patients vs. CAD. B) ROC curve of miR-149, non-CAD patients vs. unstable CAD patients. C) ROC curve of miR-149, CAD vs. unstable CAD patients. D) ROC curve of miR-424, non-CAD patients vs. CAD. E) ROC curve of miR-424, non-CAD patients vs. unstable CAD patients. F) ROC curve of miR-424, CAD vs. unstable CAD patients. G) ROC curve of miR-765, non-CAD patients vs. CAD. H) ROC curve of miR-765, non-CAD patients vs. unstable CAD patients. I) ROC curve of miR-765, CAD vs. unstable CAD patients.
Figure Legend Snippet: Diagnostic accuracy of circulating miR-149, miR-424 and miR-765 were analyzed by ROC curve. A) ROC curve of miR-149, non-CAD patients vs. CAD. B) ROC curve of miR-149, non-CAD patients vs. unstable CAD patients. C) ROC curve of miR-149, CAD vs. unstable CAD patients. D) ROC curve of miR-424, non-CAD patients vs. CAD. E) ROC curve of miR-424, non-CAD patients vs. unstable CAD patients. F) ROC curve of miR-424, CAD vs. unstable CAD patients. G) ROC curve of miR-765, non-CAD patients vs. CAD. H) ROC curve of miR-765, non-CAD patients vs. unstable CAD patients. I) ROC curve of miR-765, CAD vs. unstable CAD patients.

Techniques Used: Diagnostic Assay

11) Product Images from "Molecular Characterization and Clinical Implications of Spindle Cells in Nasopharyngeal Carcinoma: A Novel Molecule-Morphology Model of Tumor Progression Proposed"

Article Title: Molecular Characterization and Clinical Implications of Spindle Cells in Nasopharyngeal Carcinoma: A Novel Molecule-Morphology Model of Tumor Progression Proposed

Journal: PLoS ONE

doi: 10.1371/journal.pone.0083135

Neoplastic spindle cells exhibit properties of stem cells. (A) Stem cells-like markers SOX2, OCT4, Nanog and ALDH1 were markedly expressed in neoplastic spindle cells compared with neoplastic non-spindle cells (DNKC) in NPC. (B) Different expression levels of SOX2, OCT4, Nanog and ALDH1 proteins in human NPC C666-1 (undifferentiated, spindle-like morphology) and CNE1 (well differentiated, epithelial-like morphology) cell lines detected by immunofluorescence analysis.
Figure Legend Snippet: Neoplastic spindle cells exhibit properties of stem cells. (A) Stem cells-like markers SOX2, OCT4, Nanog and ALDH1 were markedly expressed in neoplastic spindle cells compared with neoplastic non-spindle cells (DNKC) in NPC. (B) Different expression levels of SOX2, OCT4, Nanog and ALDH1 proteins in human NPC C666-1 (undifferentiated, spindle-like morphology) and CNE1 (well differentiated, epithelial-like morphology) cell lines detected by immunofluorescence analysis.

Techniques Used: Expressing, Immunofluorescence

Stem-like spindle cells are strongly associated with EMT -like phenotypic changes and EBV infection. (A) Serial sections from the same patient showed aberrant expression of EMT indicators E-cahderin, N-cadherin and Vimentin in stem-like spindle cells. (B) Spindle-like C666-1 cells had strong fluorescence staining of N-cadherin and Vimentin, and weak staining of E-cadherin compared with epithelial-like CNE-1 cells. (C) Representative images show that high expression (dark brown) of EBV-encoded small RNA (EBER) was observed in neoplastic spindle cells in comparison with non-spindle cells in the same tumor. (D) The significant relationship was found between EBER, EBV-encoded protein LMP1 and CSCs/EMT-related markers. Differences statistically significant at P values
Figure Legend Snippet: Stem-like spindle cells are strongly associated with EMT -like phenotypic changes and EBV infection. (A) Serial sections from the same patient showed aberrant expression of EMT indicators E-cahderin, N-cadherin and Vimentin in stem-like spindle cells. (B) Spindle-like C666-1 cells had strong fluorescence staining of N-cadherin and Vimentin, and weak staining of E-cadherin compared with epithelial-like CNE-1 cells. (C) Representative images show that high expression (dark brown) of EBV-encoded small RNA (EBER) was observed in neoplastic spindle cells in comparison with non-spindle cells in the same tumor. (D) The significant relationship was found between EBER, EBV-encoded protein LMP1 and CSCs/EMT-related markers. Differences statistically significant at P values

Techniques Used: Infection, Expressing, Fluorescence, Staining

12) Product Images from "Molecular Characterization and Clinical Implications of Spindle Cells in Nasopharyngeal Carcinoma: A Novel Molecule-Morphology Model of Tumor Progression Proposed"

Article Title: Molecular Characterization and Clinical Implications of Spindle Cells in Nasopharyngeal Carcinoma: A Novel Molecule-Morphology Model of Tumor Progression Proposed

Journal: PLoS ONE

doi: 10.1371/journal.pone.0083135

Neoplastic spindle cells exhibit properties of stem cells. (A) Stem cells-like markers SOX2, OCT4, Nanog and ALDH1 were markedly expressed in neoplastic spindle cells compared with neoplastic non-spindle cells (DNKC) in NPC. (B) Different expression levels of SOX2, OCT4, Nanog and ALDH1 proteins in human NPC C666-1 (undifferentiated, spindle-like morphology) and CNE1 (well differentiated, epithelial-like morphology) cell lines detected by immunofluorescence analysis.
Figure Legend Snippet: Neoplastic spindle cells exhibit properties of stem cells. (A) Stem cells-like markers SOX2, OCT4, Nanog and ALDH1 were markedly expressed in neoplastic spindle cells compared with neoplastic non-spindle cells (DNKC) in NPC. (B) Different expression levels of SOX2, OCT4, Nanog and ALDH1 proteins in human NPC C666-1 (undifferentiated, spindle-like morphology) and CNE1 (well differentiated, epithelial-like morphology) cell lines detected by immunofluorescence analysis.

Techniques Used: Expressing, Immunofluorescence

13) Product Images from "Contribution of Eukaryotic-Type Serine/Threonine Kinase to Stress Response and Virulence of Streptococcus suis"

Article Title: Contribution of Eukaryotic-Type Serine/Threonine Kinase to Stress Response and Virulence of Streptococcus suis

Journal: PLoS ONE

doi: 10.1371/journal.pone.0091971

Virulence gene expression of different strains in vitro . Total RNA was extracted from SS2-1, Δ stk and C Δ stk grown in THY medium at an OD600 of 0.6–0.8 and used for qRT-PCR analysis. The mRNA level of each gene was normalized to that of 16S rRNA . Results are shown as relative expression ratios compared to expression in the parental strain SS2-1. Data from three independent assays are presented as the means±SE. Differences between SS2-1 and Δ stk was determined by two-way ANOVA. (**p
Figure Legend Snippet: Virulence gene expression of different strains in vitro . Total RNA was extracted from SS2-1, Δ stk and C Δ stk grown in THY medium at an OD600 of 0.6–0.8 and used for qRT-PCR analysis. The mRNA level of each gene was normalized to that of 16S rRNA . Results are shown as relative expression ratios compared to expression in the parental strain SS2-1. Data from three independent assays are presented as the means±SE. Differences between SS2-1 and Δ stk was determined by two-way ANOVA. (**p

Techniques Used: Expressing, In Vitro, Quantitative RT-PCR

14) Product Images from "The long non-coding RNA HNF1A-AS1 regulates proliferation and metastasis in lung adenocarcinoma"

Article Title: The long non-coding RNA HNF1A-AS1 regulates proliferation and metastasis in lung adenocarcinoma

Journal: Oncotarget

doi:

HNF1A-AS1 expression in non-small cell lung cancer (NSCLC) cells (A) qRT-PCR results demonstrating HNF1A-AS1 expression in NSCLC cell lines (A549, SPC-A1, SK-MES-1, H1703, H520) compared to human bronchial epithelial cells (16HBE). (B) HNF1A-AS1 expression, measured by qRT-PCR, following the treatment of A549 and SPC-A1 cells with scramble or with si-RNA HNF1A-AS1. (C) HNF1A-AS1 nuclear localization, as identified using qRT-PCR in fractionated A549 and SPC-A1 cells. GAPDH was used as a cytosol marker and U1 was used as a nucleus marker. * p
Figure Legend Snippet: HNF1A-AS1 expression in non-small cell lung cancer (NSCLC) cells (A) qRT-PCR results demonstrating HNF1A-AS1 expression in NSCLC cell lines (A549, SPC-A1, SK-MES-1, H1703, H520) compared to human bronchial epithelial cells (16HBE). (B) HNF1A-AS1 expression, measured by qRT-PCR, following the treatment of A549 and SPC-A1 cells with scramble or with si-RNA HNF1A-AS1. (C) HNF1A-AS1 nuclear localization, as identified using qRT-PCR in fractionated A549 and SPC-A1 cells. GAPDH was used as a cytosol marker and U1 was used as a nucleus marker. * p

Techniques Used: Expressing, Quantitative RT-PCR, Marker

DNMT1 binds to HNF1A-AS1 HNF1A-AS1 influences EMT and Cyclin D1 in lung adenocarcinoma (A) Bioinformatics analysis showed HNF1A-AS1 was enriched in sequencing data of DNMT1 RIP-seq data (GSE32260) (B) RNA immunoprecipitation (RIP) experiments were performed in A549 using the anti-DNMT1 antibody and the coprecipitated RNA was subjected to qRT-PCR for HNF1A-AS1. RIP enrichment was determined as RNA associated with DNMT1 relative to the input control. (C) CpG islands were found in the E-cadherin promoter regions by Methyl Primer Express Software. ChIP–qPCR of DNMT1 binding to the E-cadherin promoter in A549 cells (D) Gene set enrichment analysis (GSEA) indicated that HNF1A-AS1 expression was associated with gene sets involved in metastasis, microenvironment and Cyclin D1. (data downloaded from http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE48240 ) (E) Analysis of cyclinD1, E-cadherin N-cadherin and β-catenin expression in A549 and SPC-A1 cells treated with siRNA HNF1A-AS1 by western blot. (F) Analysis of β-catenin expression in A549 cells treated with siRNA HNF1A-AS1 by immunofluorescence. * p
Figure Legend Snippet: DNMT1 binds to HNF1A-AS1 HNF1A-AS1 influences EMT and Cyclin D1 in lung adenocarcinoma (A) Bioinformatics analysis showed HNF1A-AS1 was enriched in sequencing data of DNMT1 RIP-seq data (GSE32260) (B) RNA immunoprecipitation (RIP) experiments were performed in A549 using the anti-DNMT1 antibody and the coprecipitated RNA was subjected to qRT-PCR for HNF1A-AS1. RIP enrichment was determined as RNA associated with DNMT1 relative to the input control. (C) CpG islands were found in the E-cadherin promoter regions by Methyl Primer Express Software. ChIP–qPCR of DNMT1 binding to the E-cadherin promoter in A549 cells (D) Gene set enrichment analysis (GSEA) indicated that HNF1A-AS1 expression was associated with gene sets involved in metastasis, microenvironment and Cyclin D1. (data downloaded from http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE48240 ) (E) Analysis of cyclinD1, E-cadherin N-cadherin and β-catenin expression in A549 and SPC-A1 cells treated with siRNA HNF1A-AS1 by western blot. (F) Analysis of β-catenin expression in A549 cells treated with siRNA HNF1A-AS1 by immunofluorescence. * p

Techniques Used: Sequencing, Immunoprecipitation, Quantitative RT-PCR, Software, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Binding Assay, Expressing, Western Blot, Immunofluorescence

15) Product Images from "Wnt3a upregulates brain-derived insulin by increasing NeuroD1 via Wnt/β-catenin signaling in the hypothalamus"

Article Title: Wnt3a upregulates brain-derived insulin by increasing NeuroD1 via Wnt/β-catenin signaling in the hypothalamus

Journal: Molecular Brain

doi: 10.1186/s13041-016-0207-5

Wnt3a-induced Ins2 upregulation is blocked by knockdown of NeuroD1. NeuroD1 was knocked down by infecting N39 cells with lentivirus containing shRNA for 72 h. a N39 cells were infected by lentiviral shRNA against NeuroD1 for 48 h and then treated with Wnt3a (100 ng/mL) for 24 h. The levels of active and total β-catenin and NeuroD1 protein were determined by immunoblot assay. b The intensity of NeuroD1 signal in ( a ) was quantified using the ImageJ software ( n = 4). The levels of NeuroD1 mRNA ( c ) and Ins2 mRNA ( d ) were measured by qRT-PCR in N39 cells that were infected with lentiviral non-targeting or NeuroD1 shRNA for 48 h and then treated with Wnt3a (100 ng/mL) for 24 h ( n = 9). Data are means + SEM. *** p
Figure Legend Snippet: Wnt3a-induced Ins2 upregulation is blocked by knockdown of NeuroD1. NeuroD1 was knocked down by infecting N39 cells with lentivirus containing shRNA for 72 h. a N39 cells were infected by lentiviral shRNA against NeuroD1 for 48 h and then treated with Wnt3a (100 ng/mL) for 24 h. The levels of active and total β-catenin and NeuroD1 protein were determined by immunoblot assay. b The intensity of NeuroD1 signal in ( a ) was quantified using the ImageJ software ( n = 4). The levels of NeuroD1 mRNA ( c ) and Ins2 mRNA ( d ) were measured by qRT-PCR in N39 cells that were infected with lentiviral non-targeting or NeuroD1 shRNA for 48 h and then treated with Wnt3a (100 ng/mL) for 24 h ( n = 9). Data are means + SEM. *** p

Techniques Used: shRNA, Infection, Software, Quantitative RT-PCR

Wnt3a upregulates NeuroD1 and Ins2 in the mouse hypothalamus. The basal mRNA levels of Ins2 and Wnt3a were measured by qRT-PCR in hypothalamic tissues from C57BL/6 male mice ( n = 6−9) ( a ). C57BL/6 male mice received icv injections of vehicle or Wnt3a (4 or 20 ng), and hypothalamic tissues were dissected after 24 h. The mRNA levels of Ins2 ( b ) and NeuroD1 ( c ) were measured by qRT-PCR ( n = 6−9). Data are means + SEM. ### p
Figure Legend Snippet: Wnt3a upregulates NeuroD1 and Ins2 in the mouse hypothalamus. The basal mRNA levels of Ins2 and Wnt3a were measured by qRT-PCR in hypothalamic tissues from C57BL/6 male mice ( n = 6−9) ( a ). C57BL/6 male mice received icv injections of vehicle or Wnt3a (4 or 20 ng), and hypothalamic tissues were dissected after 24 h. The mRNA levels of Ins2 ( b ) and NeuroD1 ( c ) were measured by qRT-PCR ( n = 6−9). Data are means + SEM. ### p

Techniques Used: Quantitative RT-PCR, Mouse Assay

The proposed regulatory mechanism of insulin expression by the Wnt/β-catenin/NeuroD1 pathway in the hypothalamus. Wnt3a might bind to LRP and Frizzed receptor to activate the canonical Wnt pathway. GSK3 phosphorylates β-catenin and induces its degradation; Wnt3a inhibits GSK3, leading to β-catenin accumulation. Accumulated β-catenin translocates into the nucleus and enhances the expression of NeuroD1 with TCF. Up-regulated NeuroD1 also translocates into the nucleus, where it induces the production of insulin
Figure Legend Snippet: The proposed regulatory mechanism of insulin expression by the Wnt/β-catenin/NeuroD1 pathway in the hypothalamus. Wnt3a might bind to LRP and Frizzed receptor to activate the canonical Wnt pathway. GSK3 phosphorylates β-catenin and induces its degradation; Wnt3a inhibits GSK3, leading to β-catenin accumulation. Accumulated β-catenin translocates into the nucleus and enhances the expression of NeuroD1 with TCF. Up-regulated NeuroD1 also translocates into the nucleus, where it induces the production of insulin

Techniques Used: Expressing

Knockdown of NeuroD1 reduces the expression of Ins2 . NeuroD1 was knocked down by infecting N39 cells with lentivirus containing shRNA (shNeuroD1) for 72 h. a Knockdown efficiency was determined by examining the levels of NeuroD1 mRNA using qRT-PCR ( n = 23). b The level of NeuroD1 protein was assessed by immunoblot assay, and c the intensity of NeuroD1 bands was quantified using the ImageJ software with normalization to GAPDH ( n = 6). d The basal expression of Ins2 was determined by qRT-PCR after infection with shRNA lentivirus against NeuroD1 for 72 h ( n = 12). Data are means + SEM. ** p
Figure Legend Snippet: Knockdown of NeuroD1 reduces the expression of Ins2 . NeuroD1 was knocked down by infecting N39 cells with lentivirus containing shRNA (shNeuroD1) for 72 h. a Knockdown efficiency was determined by examining the levels of NeuroD1 mRNA using qRT-PCR ( n = 23). b The level of NeuroD1 protein was assessed by immunoblot assay, and c the intensity of NeuroD1 bands was quantified using the ImageJ software with normalization to GAPDH ( n = 6). d The basal expression of Ins2 was determined by qRT-PCR after infection with shRNA lentivirus against NeuroD1 for 72 h ( n = 12). Data are means + SEM. ** p

Techniques Used: Expressing, shRNA, Quantitative RT-PCR, Software, Infection

Wnt3a increases the expression of NeuroD1 by activating Wnt/β-catenin signaling in N39 cells. a NeuroD1 mRNA levels were quantified using qRT-PCR after vehicle or Wnt3a (25 or 100 ng/mL) treatment for 6, 12, and 24 h ( n = 23). b NeuroD1 protein levels were measured by immunoblot assay after vehicle or Wnt3a (20 or 100 ng/mL) treatment for 12 and 24 h. c The intensity of bands shown in ( b ) was quantified by using ImageJ with normalization to GAPDH ( n = 6). d After treatment with vehicle or Wnt3a (100 ng/mL) for 24 h, NeuroD1 was examined using the immunofluorescence assay; the nuclei were stained with Hoechst 33342 dye. Scale bar, 20 μm. e The level of NeuroD1 mRNA was determined by qRT-PCR 12 h after treatment with 1 μM BIO ( n = 9). f Cells were treated with 1 μM BIO, and the level of NeuroD1 was measured by immunoblot assay after treatment for 1, 3, 6, and 12 h. g The intensity of NeuroD1 bands in ( f ) was quantified using the ImageJ software and normalized to GAPDH ( n = 6). Data are means + SEM. * p
Figure Legend Snippet: Wnt3a increases the expression of NeuroD1 by activating Wnt/β-catenin signaling in N39 cells. a NeuroD1 mRNA levels were quantified using qRT-PCR after vehicle or Wnt3a (25 or 100 ng/mL) treatment for 6, 12, and 24 h ( n = 23). b NeuroD1 protein levels were measured by immunoblot assay after vehicle or Wnt3a (20 or 100 ng/mL) treatment for 12 and 24 h. c The intensity of bands shown in ( b ) was quantified by using ImageJ with normalization to GAPDH ( n = 6). d After treatment with vehicle or Wnt3a (100 ng/mL) for 24 h, NeuroD1 was examined using the immunofluorescence assay; the nuclei were stained with Hoechst 33342 dye. Scale bar, 20 μm. e The level of NeuroD1 mRNA was determined by qRT-PCR 12 h after treatment with 1 μM BIO ( n = 9). f Cells were treated with 1 μM BIO, and the level of NeuroD1 was measured by immunoblot assay after treatment for 1, 3, 6, and 12 h. g The intensity of NeuroD1 bands in ( f ) was quantified using the ImageJ software and normalized to GAPDH ( n = 6). Data are means + SEM. * p

Techniques Used: Expressing, Quantitative RT-PCR, Immunofluorescence, Staining, Software

16) Product Images from "Roles of Long Non-Coding RNA CCAT2 in Cervical Cancer Cell Growth and Apoptosis"

Article Title: Roles of Long Non-Coding RNA CCAT2 in Cervical Cancer Cell Growth and Apoptosis

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

doi: 10.12659/MSM.897754

Relative expression of CCAT2 in 3 cervical cancer cell lines was carried out by qRT-PCR ( A ). Knockdown effects of siRNA CCAT2 in HeLa cell line ( B ). CCK8 assay to test the function of lncRNA CCAT2. Downregulation of CCAT2 inhibited HeLa cells growth ( C ). Statistical analyses were performed with the independent samples t test. All data are shown as the mean ±SD.
Figure Legend Snippet: Relative expression of CCAT2 in 3 cervical cancer cell lines was carried out by qRT-PCR ( A ). Knockdown effects of siRNA CCAT2 in HeLa cell line ( B ). CCK8 assay to test the function of lncRNA CCAT2. Downregulation of CCAT2 inhibited HeLa cells growth ( C ). Statistical analyses were performed with the independent samples t test. All data are shown as the mean ±SD.

Techniques Used: Expressing, Quantitative RT-PCR, CCK-8 Assay

17) Product Images from "Overexpression of Poplar PtrWRKY89 in Transgenic Arabidopsis Leads to a Reduction of Disease Resistance by Regulating Defense-Related Genes in Salicylate- and Jasmonate-Dependent Signaling"

Article Title: Overexpression of Poplar PtrWRKY89 in Transgenic Arabidopsis Leads to a Reduction of Disease Resistance by Regulating Defense-Related Genes in Salicylate- and Jasmonate-Dependent Signaling

Journal: PLoS ONE

doi: 10.1371/journal.pone.0149137

PtrWRKY89 overexpressing Arabidopsis plants showed more susceptible to Pst DC3000 than wild-type plants. (A) Disease symptoms of wild-type (WT) and transgenic plants after 3 days of Pst DC3000 infection. (B) The quantification of total chlorophyll content in transgenic and WT plants at 3 days post infection. (C) Growth of Pst DC3000 in planta 3 days after inoculation. Values represent means of three replicates. Error bars indicate standard deviation. Asterisks indicate a statistically significant difference between WT and transgenic plants (*, P
Figure Legend Snippet: PtrWRKY89 overexpressing Arabidopsis plants showed more susceptible to Pst DC3000 than wild-type plants. (A) Disease symptoms of wild-type (WT) and transgenic plants after 3 days of Pst DC3000 infection. (B) The quantification of total chlorophyll content in transgenic and WT plants at 3 days post infection. (C) Growth of Pst DC3000 in planta 3 days after inoculation. Values represent means of three replicates. Error bars indicate standard deviation. Asterisks indicate a statistically significant difference between WT and transgenic plants (*, P

Techniques Used: Transgenic Assay, Infection, Standard Deviation

Expression analysis of the PtrWRKY89 promoter. (A) The promoter of PtrWRKY89 was cloned and ligated into vector pCXGUS-P to drive GUS expression and the resulting construct was introduced into A . thaliana . Transgenic seedlings were grown on MS media and then transplanted in soil for GUS staining. GUS expression was observed in various tissues of transgenic plants, including (B) flowers, (C) siliques, (D) 5-week-old seedlings, (E) old leaves, (F) young leaves and (G) roots.
Figure Legend Snippet: Expression analysis of the PtrWRKY89 promoter. (A) The promoter of PtrWRKY89 was cloned and ligated into vector pCXGUS-P to drive GUS expression and the resulting construct was introduced into A . thaliana . Transgenic seedlings were grown on MS media and then transplanted in soil for GUS staining. GUS expression was observed in various tissues of transgenic plants, including (B) flowers, (C) siliques, (D) 5-week-old seedlings, (E) old leaves, (F) young leaves and (G) roots.

Techniques Used: Expressing, Clone Assay, Plasmid Preparation, Construct, Transgenic Assay, Mass Spectrometry, Staining

Constitutive expressing PtrWRKY89 in Arabidopsis plants showing susceptibility to Botrytis cinerea . (A) Disease response of inoculated plants at 7 days after B . cinerea infection. (B) Transcript accumulation of B . cinerea Actin gene in these inoculated plants. Arabidopsis UBC gene was used as an internal control. Values represent means of three replicates and error bars indicated standard deviation. Two asterisks indicated a statistically significant difference between WT and transgenic plants (**, P
Figure Legend Snippet: Constitutive expressing PtrWRKY89 in Arabidopsis plants showing susceptibility to Botrytis cinerea . (A) Disease response of inoculated plants at 7 days after B . cinerea infection. (B) Transcript accumulation of B . cinerea Actin gene in these inoculated plants. Arabidopsis UBC gene was used as an internal control. Values represent means of three replicates and error bars indicated standard deviation. Two asterisks indicated a statistically significant difference between WT and transgenic plants (**, P

Techniques Used: Expressing, Infection, Standard Deviation, Transgenic Assay

Gene expression analyses of JA-signaling pathway genes in transgenic Arabidopsis overexpressed PtrWRKY89 after spraying spore suspending of B . cinerea . AOS : ALLENE OXIDE SYNTHASE , JAR1 : JASMONATE RESISTANT 1 , JAZ1/JAZ2/JAZ9/JAZ10 : JASMONATE-ZIM-DOMAIN PROTEIN 1/2/9/10 , COI1 : CORONATINE INSENSITIVE 1 , ERF1 : ETHYLENE RESPONSE FACTOR 1 , ORA59 : OCTADECANOID-RESPONSIVE ARABIDOPSIS AP2/ERF 59 , PDF1 . 2 : PLANT DEFENSIN 1 . 2 . Arabidopsis UBC gene was used as an internal control.
Figure Legend Snippet: Gene expression analyses of JA-signaling pathway genes in transgenic Arabidopsis overexpressed PtrWRKY89 after spraying spore suspending of B . cinerea . AOS : ALLENE OXIDE SYNTHASE , JAR1 : JASMONATE RESISTANT 1 , JAZ1/JAZ2/JAZ9/JAZ10 : JASMONATE-ZIM-DOMAIN PROTEIN 1/2/9/10 , COI1 : CORONATINE INSENSITIVE 1 , ERF1 : ETHYLENE RESPONSE FACTOR 1 , ORA59 : OCTADECANOID-RESPONSIVE ARABIDOPSIS AP2/ERF 59 , PDF1 . 2 : PLANT DEFENSIN 1 . 2 . Arabidopsis UBC gene was used as an internal control.

Techniques Used: Expressing, Transgenic Assay

Expression of SA-related genes in transgenic Arabidopsis overexpressed PtrWRKY89 after inoculation of Pst DC3000. ICS1 : ISOCHORISMATE SYNTHASE 1 , PAD4 : PHYTOALEXIN DEFICIENT 4 , EDS1 : ENHANCED DISEASE SUSCEPTIBILITY 1 , MYB44 : ARABIDOPSIS THALIANA MYB DOMAIN PROTEIN 44 , NPR1 : NONEXPRESSER OF PR GENES 1 , CPR5 : CONSTITUTIVE EXPRESSION OF PR GENES 5 , PR1/PR2/PR5 : PATHOGENESIS RELATED GENES 1/2/5 . Arabidopsis UBC gene was used as an internal control.
Figure Legend Snippet: Expression of SA-related genes in transgenic Arabidopsis overexpressed PtrWRKY89 after inoculation of Pst DC3000. ICS1 : ISOCHORISMATE SYNTHASE 1 , PAD4 : PHYTOALEXIN DEFICIENT 4 , EDS1 : ENHANCED DISEASE SUSCEPTIBILITY 1 , MYB44 : ARABIDOPSIS THALIANA MYB DOMAIN PROTEIN 44 , NPR1 : NONEXPRESSER OF PR GENES 1 , CPR5 : CONSTITUTIVE EXPRESSION OF PR GENES 5 , PR1/PR2/PR5 : PATHOGENESIS RELATED GENES 1/2/5 . Arabidopsis UBC gene was used as an internal control.

Techniques Used: Expressing, Transgenic Assay

Subcellular localization and transactivation assay of PtrWRKY89. (A) PtrWRKY89 was ligated into pCX-DG vector to generate GFP : PtrWRKY89 construct. The resulting construct and empty vectors were transformed into epidermal cells of onion ( Allium cepa ) and stained with DAPI, respectively. The fusion protein displayed its localization to the cell nucleus as manifested by GFP (lower column) and GFP driven by the CaMV 35 promoter was localized to both the cytoplasm and the cell nucleus (upper column). Overlay and bright field images of the epidermal cells were also shown. (B) PtrWRKY89 was cloned into pGBKT7 vector with DNA binding domain of GAL4 and introduced into Gold2 yeast cells. The transformants were grown on SD medium lacking tryptophan (Try) for the sake of positive clone selection and then on SD medium without Try, histidine (His) and adenine (Ade) for the transactivation assay. The clones grown on SD (-Try/His/Ade) were stained by X-α-gal. GAL4-BD (empty vector) was a negative control.
Figure Legend Snippet: Subcellular localization and transactivation assay of PtrWRKY89. (A) PtrWRKY89 was ligated into pCX-DG vector to generate GFP : PtrWRKY89 construct. The resulting construct and empty vectors were transformed into epidermal cells of onion ( Allium cepa ) and stained with DAPI, respectively. The fusion protein displayed its localization to the cell nucleus as manifested by GFP (lower column) and GFP driven by the CaMV 35 promoter was localized to both the cytoplasm and the cell nucleus (upper column). Overlay and bright field images of the epidermal cells were also shown. (B) PtrWRKY89 was cloned into pGBKT7 vector with DNA binding domain of GAL4 and introduced into Gold2 yeast cells. The transformants were grown on SD medium lacking tryptophan (Try) for the sake of positive clone selection and then on SD medium without Try, histidine (His) and adenine (Ade) for the transactivation assay. The clones grown on SD (-Try/His/Ade) were stained by X-α-gal. GAL4-BD (empty vector) was a negative control.

Techniques Used: Transactivation Assay, Plasmid Preparation, Construct, Transformation Assay, Staining, Clone Assay, Binding Assay, Selection, Negative Control

Expression patterns of the PtrWRKY89 promoter driven GUS gene in response to salicylic acid (SA), methyl jasmonate (MeJA) with low concentration and their mixed solution, respectively. SA solution (100 μΜ), MeJA solution (100 μΜ) and SA+MeJA mixture (100 μΜ + 100 μΜ) were sprayed onto the surface of two-week-old transgenic Arabidopsis seedling containing the ProPtrWRKY89-GUS construct. The control plants (CK) were treated by H 2 O. After treatments of 24 h, the seedlings were collected from MS medium. (A) The GUS staining of the seedlings. (B) The transcript profiles of PtrWRKY89 were analyzed by quantitative RT-PCR (qRT-PCR). Error bars were obtained from three biological replicates. Arabidopsis UBC (AT5G25760) expression was used as a control and gene-specific primers were used for qRT-PCR analysis were described in S1 Table . Two asterisks indicate a statistically significant difference by Student's t -test (**, P
Figure Legend Snippet: Expression patterns of the PtrWRKY89 promoter driven GUS gene in response to salicylic acid (SA), methyl jasmonate (MeJA) with low concentration and their mixed solution, respectively. SA solution (100 μΜ), MeJA solution (100 μΜ) and SA+MeJA mixture (100 μΜ + 100 μΜ) were sprayed onto the surface of two-week-old transgenic Arabidopsis seedling containing the ProPtrWRKY89-GUS construct. The control plants (CK) were treated by H 2 O. After treatments of 24 h, the seedlings were collected from MS medium. (A) The GUS staining of the seedlings. (B) The transcript profiles of PtrWRKY89 were analyzed by quantitative RT-PCR (qRT-PCR). Error bars were obtained from three biological replicates. Arabidopsis UBC (AT5G25760) expression was used as a control and gene-specific primers were used for qRT-PCR analysis were described in S1 Table . Two asterisks indicate a statistically significant difference by Student's t -test (**, P

Techniques Used: Expressing, Concentration Assay, Transgenic Assay, Construct, Mass Spectrometry, Staining, Quantitative RT-PCR

18) Product Images from "Comprehensive Identification and Bread-Making Quality Evaluation of Common Wheat Somatic Variation Line AS208 on Glutenin Composition"

Article Title: Comprehensive Identification and Bread-Making Quality Evaluation of Common Wheat Somatic Variation Line AS208 on Glutenin Composition

Journal: PLoS ONE

doi: 10.1371/journal.pone.0146933

Expression profile of the 1Bx20 gene at different development stages in LX987 and AS208 assessed by qRT-PCR. The 1Bx20 gene was not expressed at any stages during grain development in AS208 (A8-A29). Its expression in LX987 started from the 11 th day since anthesis (L11), reached its highest level at the 19 th day post anthesis, and then gradually declined as filling progressed (L21-L29).
Figure Legend Snippet: Expression profile of the 1Bx20 gene at different development stages in LX987 and AS208 assessed by qRT-PCR. The 1Bx20 gene was not expressed at any stages during grain development in AS208 (A8-A29). Its expression in LX987 started from the 11 th day since anthesis (L11), reached its highest level at the 19 th day post anthesis, and then gradually declined as filling progressed (L21-L29).

Techniques Used: Expressing, Quantitative RT-PCR

Identification of seed storage proteins in AS208, LX987, and CS by RP-HPLC. The peaks for albumin (a), globulin (b), and gliadin (c) in AS208 and LX987 were very similar, but were different from those of CS. However, the peak patterns for glutenin (d) were different among AS208, LX987, and CS, in which the three wheat accessions all had peaks for 1Dx2 and 1Dy12, while AS208 was missing peaks for 1Bx20 and 1By20 compared to its progenitor genotype LX987. The grains were harvested in Beijing in 2013.
Figure Legend Snippet: Identification of seed storage proteins in AS208, LX987, and CS by RP-HPLC. The peaks for albumin (a), globulin (b), and gliadin (c) in AS208 and LX987 were very similar, but were different from those of CS. However, the peak patterns for glutenin (d) were different among AS208, LX987, and CS, in which the three wheat accessions all had peaks for 1Dx2 and 1Dy12, while AS208 was missing peaks for 1Bx20 and 1By20 compared to its progenitor genotype LX987. The grains were harvested in Beijing in 2013.

Techniques Used: High Performance Liquid Chromatography

19) Product Images from "Transcription factors Asg1p and Hal9p regulate pH homeostasis in Candida glabrata"

Article Title: Transcription factors Asg1p and Hal9p regulate pH homeostasis in Candida glabrata

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2015.00843

An interaction between CgASG1 and CgHAL9 was tested using quantitative reverse-transcribed PCR (qRT-PCR) and green fluorescent protein (GFP) localization. (A) qRT-PCR was performed on RNA samples of log-phase cells in YNB and YNB-pH 2.0 media to analyze the expression levels of CgASG1 and CgHAL9 in the indicated strains. Results are presented as fold expression relative to the levels of the wild-type β-actin control at 0 and 2-h time points of incubation in YNB and YNB-pH 2.0 media. The means and standard deviations for three independent experiments are shown. (B) Log-phase Cgasg1 Δ/ CgHAL9 - GFP and Cghal9 Δ/ CgASG1 - GFP cells were grown in YNB-pH 2.0 medium for 2 h, and the localization of GFP fusion proteins was determined by fluorescence microscopy.
Figure Legend Snippet: An interaction between CgASG1 and CgHAL9 was tested using quantitative reverse-transcribed PCR (qRT-PCR) and green fluorescent protein (GFP) localization. (A) qRT-PCR was performed on RNA samples of log-phase cells in YNB and YNB-pH 2.0 media to analyze the expression levels of CgASG1 and CgHAL9 in the indicated strains. Results are presented as fold expression relative to the levels of the wild-type β-actin control at 0 and 2-h time points of incubation in YNB and YNB-pH 2.0 media. The means and standard deviations for three independent experiments are shown. (B) Log-phase Cgasg1 Δ/ CgHAL9 - GFP and Cghal9 Δ/ CgASG1 - GFP cells were grown in YNB-pH 2.0 medium for 2 h, and the localization of GFP fusion proteins was determined by fluorescence microscopy.

Techniques Used: Polymerase Chain Reaction, Quantitative RT-PCR, Expressing, Incubation, Fluorescence, Microscopy

Quantitative reverse-transcribed PCR (qRT-PCR) validation of transcriptional profiles in an acidic environment . Logarithmic-phase C. glabrata cells were incubated in YNB or YNB-pH 2.0 media for 2 h. qRT-PCR analyses of the indicated genes was performed as described in the Materials and Methods Section. The means and standard deviations for three independent experiments are shown. C. glabrata strains: wild-type, Cgasg1 Δ and Cghal9 Δ strains. ( * P
Figure Legend Snippet: Quantitative reverse-transcribed PCR (qRT-PCR) validation of transcriptional profiles in an acidic environment . Logarithmic-phase C. glabrata cells were incubated in YNB or YNB-pH 2.0 media for 2 h. qRT-PCR analyses of the indicated genes was performed as described in the Materials and Methods Section. The means and standard deviations for three independent experiments are shown. C. glabrata strains: wild-type, Cgasg1 Δ and Cghal9 Δ strains. ( * P

Techniques Used: Polymerase Chain Reaction, Quantitative RT-PCR, Incubation

Growth assays in different YNB media. (A) Deletion of CgASG1 has no effect on the utilization of non-fermentative carbon sources in C. glabrata . (B) CgAsg1p plays a role under acid-stress conditions. (C) CgHAL9 serves an important role in cell growth under hypersaline conditions in C. glabrata . (D) CgHal9p plays a role under acid-stress conditions. Logarithmic-phase cells of each C. glabrata strain were adjusted to 2 × 10 7 cells/mL, and then 4 μL of serial tenfold dilutions were spotted onto the corresponding YNB media, as indicated. Pictures were taken after 4 days of growth at 30°C.
Figure Legend Snippet: Growth assays in different YNB media. (A) Deletion of CgASG1 has no effect on the utilization of non-fermentative carbon sources in C. glabrata . (B) CgAsg1p plays a role under acid-stress conditions. (C) CgHAL9 serves an important role in cell growth under hypersaline conditions in C. glabrata . (D) CgHal9p plays a role under acid-stress conditions. Logarithmic-phase cells of each C. glabrata strain were adjusted to 2 × 10 7 cells/mL, and then 4 μL of serial tenfold dilutions were spotted onto the corresponding YNB media, as indicated. Pictures were taken after 4 days of growth at 30°C.

Techniques Used:

20) Product Images from "MicroRNA-146a-5p Negatively Regulates Pro-Inflammatory Cytokine Secretion and Cell Activation in Lipopolysaccharide Stimulated Human Hepatic Stellate Cells through Inhibition of Toll-Like Receptor 4 Signaling Pathways"

Article Title: MicroRNA-146a-5p Negatively Regulates Pro-Inflammatory Cytokine Secretion and Cell Activation in Lipopolysaccharide Stimulated Human Hepatic Stellate Cells through Inhibition of Toll-Like Receptor 4 Signaling Pathways

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms17071076

Lipopolysaccharide (LPS) up-regulated toll-like receptor 4 (TLR4) and α-smooth muscle actin (α-SMA) expression and down-regulated miR-146a-5p expression: ( A , B ) LX2 cells were treated with different amounts of LPS (0~1000 ng/mL) for 24 or 48 h and then quantitative real-time PCR (qRT-PCR) were used to detect the expression of TLR4 and α-SMA; ( C ) miR-146a-5p expression was examined by qRT-PCR after treatment with different amounts of LPS (0~1000 ng/mL) for 24 h; and ( D ) miR-146a-5p expression was assessed by qRT-PCR after treatment with 50 ng/mL of LPS for the various times (0, 6, 12, 24 and 48 h). * p
Figure Legend Snippet: Lipopolysaccharide (LPS) up-regulated toll-like receptor 4 (TLR4) and α-smooth muscle actin (α-SMA) expression and down-regulated miR-146a-5p expression: ( A , B ) LX2 cells were treated with different amounts of LPS (0~1000 ng/mL) for 24 or 48 h and then quantitative real-time PCR (qRT-PCR) were used to detect the expression of TLR4 and α-SMA; ( C ) miR-146a-5p expression was examined by qRT-PCR after treatment with different amounts of LPS (0~1000 ng/mL) for 24 h; and ( D ) miR-146a-5p expression was assessed by qRT-PCR after treatment with 50 ng/mL of LPS for the various times (0, 6, 12, 24 and 48 h). * p

Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Quantitative RT-PCR

TRAF6 siRNA reduced LPS induced JNK activation and α-SMA expression. After incubation with TRAF6 siRNA or IRAK1 siRNA for 48 h, LX2 cells were treated with 500 ng/mL of LPS for 24 h. ( A ) TRAF6 siRNA but not IRAK1 siRNA decreased the mRNA expression level of α-SMA; ( B ) TRAF6 siRNA but not IRAK1 siRNA attenuated the activation of JNK and Smad2 and decreased the protein expression of α-SMA. * p
Figure Legend Snippet: TRAF6 siRNA reduced LPS induced JNK activation and α-SMA expression. After incubation with TRAF6 siRNA or IRAK1 siRNA for 48 h, LX2 cells were treated with 500 ng/mL of LPS for 24 h. ( A ) TRAF6 siRNA but not IRAK1 siRNA decreased the mRNA expression level of α-SMA; ( B ) TRAF6 siRNA but not IRAK1 siRNA attenuated the activation of JNK and Smad2 and decreased the protein expression of α-SMA. * p

Techniques Used: Activation Assay, Expressing, Incubation

miR-146a-5p mimic attenuated LPS induced JNK activation and α-SMA expression. After pretreatment in the absence or presence of miR-146a-5p for 24 h, LX2 cells were stimulated with 500 ng/mL of LPS for 24 h: ( A ) Overexpression of miR-146a-5p significantly inhibited α-SMA mRNA expression; and ( B ) Western bolt showed that miR-146a-5p mimic significantly decreased LPS induced p-JNK, p-Smad2 and α-SMA expression while miR-146a-5p inhibitor showed the contrary changes. * p
Figure Legend Snippet: miR-146a-5p mimic attenuated LPS induced JNK activation and α-SMA expression. After pretreatment in the absence or presence of miR-146a-5p for 24 h, LX2 cells were stimulated with 500 ng/mL of LPS for 24 h: ( A ) Overexpression of miR-146a-5p significantly inhibited α-SMA mRNA expression; and ( B ) Western bolt showed that miR-146a-5p mimic significantly decreased LPS induced p-JNK, p-Smad2 and α-SMA expression while miR-146a-5p inhibitor showed the contrary changes. * p

Techniques Used: Activation Assay, Expressing, Over Expression, Western Blot

21) Product Images from "Virus-Induced Gene Silencing-Based Functional Analyses Revealed the Involvement of Several Putative Trehalose-6-Phosphate Synthase/Phosphatase Genes in Disease Resistance against Botrytis cinerea and Pseudomonas syringae pv. tomato DC3000 in Tomato"

Article Title: Virus-Induced Gene Silencing-Based Functional Analyses Revealed the Involvement of Several Putative Trehalose-6-Phosphate Synthase/Phosphatase Genes in Disease Resistance against Botrytis cinerea and Pseudomonas syringae pv. tomato DC3000 in Tomato

Journal: Frontiers in Plant Science

doi: 10.3389/fpls.2016.01176

Silencing efficiency and specificity for selected SlTPS , SlTPP and SlTRE1 genes in VIGS-infiltrated plants. (A) Silencing efficiency for each of the selected trehalose-related genes in corresponding VIGS-infiltrated plants. (B,C) Silencing specificity for 4 SlTPS genes and for SlTPP2 . Ten-day-old tomato plants were infiltrated with agrobacteria carrying TRV-SlTPSs/SlTPPs/SlTRE1 or TRV-GUS constructs and leaf samples were collected at 4 weeks after agroinfiltration. Transcript levels for the selected trehalose-related genes were analyzed by qRT-PCR using a tomato SlActin gene as an internal control. Expression levels of the selected trehalose-related genes in TRV-SlTPSs/SlTPPs/SlTRE1-infiltrated plants were shown as percentages of the levels in TRV-GUS-infiltrated plants. Data presented are the means ± SD from three independent experiments and ∗ above the columns indicate significant differences at p
Figure Legend Snippet: Silencing efficiency and specificity for selected SlTPS , SlTPP and SlTRE1 genes in VIGS-infiltrated plants. (A) Silencing efficiency for each of the selected trehalose-related genes in corresponding VIGS-infiltrated plants. (B,C) Silencing specificity for 4 SlTPS genes and for SlTPP2 . Ten-day-old tomato plants were infiltrated with agrobacteria carrying TRV-SlTPSs/SlTPPs/SlTRE1 or TRV-GUS constructs and leaf samples were collected at 4 weeks after agroinfiltration. Transcript levels for the selected trehalose-related genes were analyzed by qRT-PCR using a tomato SlActin gene as an internal control. Expression levels of the selected trehalose-related genes in TRV-SlTPSs/SlTPPs/SlTRE1-infiltrated plants were shown as percentages of the levels in TRV-GUS-infiltrated plants. Data presented are the means ± SD from three independent experiments and ∗ above the columns indicate significant differences at p

Techniques Used: Construct, Quantitative RT-PCR, Expressing

Silencing of SlTPS3 , SlTPS4 , and SlTPS7 led to decreased resistance against B. cinerea in whole plant assays. (A) Disease phenotype of representative TRV-SlTPS3-, TRV-SlTPS4-, TRV-SlTPS7-, and TRV-GUS-infiltrated plants. Photos were taken at 4 days after inoculation. (B) In planta growth of B. cinerea in inoculated TRV-SlTPS3-, TRV-SlTPS4-, TRV-SlTPS7-, and TRV-GUS-infiltrated plants. Whole plant disease assays were done by foliar spraying with spore suspension at 4 weeks after VIGS infiltration. Transcript levels for B. cinerea BcActinA and tomato SlActin genes in B. cinerea -inoculated plants were analyzed using qRT-PCR and in planta relative growth of B. cinerea was shown as ratios of transcript levels of BcActinA / SlActin . Similar results were obtained in independent experiments (A) and data presented in (B) are the means ± SD from three independent experiments. ∗ above the columns indicate significant differences at p
Figure Legend Snippet: Silencing of SlTPS3 , SlTPS4 , and SlTPS7 led to decreased resistance against B. cinerea in whole plant assays. (A) Disease phenotype of representative TRV-SlTPS3-, TRV-SlTPS4-, TRV-SlTPS7-, and TRV-GUS-infiltrated plants. Photos were taken at 4 days after inoculation. (B) In planta growth of B. cinerea in inoculated TRV-SlTPS3-, TRV-SlTPS4-, TRV-SlTPS7-, and TRV-GUS-infiltrated plants. Whole plant disease assays were done by foliar spraying with spore suspension at 4 weeks after VIGS infiltration. Transcript levels for B. cinerea BcActinA and tomato SlActin genes in B. cinerea -inoculated plants were analyzed using qRT-PCR and in planta relative growth of B. cinerea was shown as ratios of transcript levels of BcActinA / SlActin . Similar results were obtained in independent experiments (A) and data presented in (B) are the means ± SD from three independent experiments. ∗ above the columns indicate significant differences at p

Techniques Used: Quantitative RT-PCR

22) Product Images from "A novel hypoxia-induced miR-147a regulates cell proliferation through a positive feedback loop of stabilizing HIF-1α"

Article Title: A novel hypoxia-induced miR-147a regulates cell proliferation through a positive feedback loop of stabilizing HIF-1α

Journal: Cancer Biology & Therapy

doi: 10.1080/15384047.2016.1195040

Hypoxia-induced miR-147 inhibits cell proliferation and colony formation. (A) Hypoxia induces miR-147a expression. HeLa cells were exposed to 1% oxygen for 12, 24 and 48 h; real-time PCR analysis of miR-147a levels in HeLa cells was done and compared with normoxia (21% O 2 ). U6 small nuclear RNA was used as an internal control. Data shown are mean ± SD of 3 independent experiments. *P
Figure Legend Snippet: Hypoxia-induced miR-147 inhibits cell proliferation and colony formation. (A) Hypoxia induces miR-147a expression. HeLa cells were exposed to 1% oxygen for 12, 24 and 48 h; real-time PCR analysis of miR-147a levels in HeLa cells was done and compared with normoxia (21% O 2 ). U6 small nuclear RNA was used as an internal control. Data shown are mean ± SD of 3 independent experiments. *P

Techniques Used: Expressing, Real-time Polymerase Chain Reaction

23) Product Images from "Recognition of tumor cells by Dectin-1 orchestrates innate immune cells for anti-tumor responses"

Article Title: Recognition of tumor cells by Dectin-1 orchestrates innate immune cells for anti-tumor responses

Journal: eLife

doi: 10.7554/eLife.04177

Induction of Inam mRNAs in DCs co-cultured with human cancer cells. Dectin-1-dependent induction of Inam ( Fam26f ) mRNA by a human cancer cell line. Mouse splenic CD11c + cells (3 × 10 5 cells), either from WT mice or Dectin-1 −/− mice, were co-cultured with HBC4 cells (1 × 10 4 cells) for 4 hr and total RNA was subjected to qRT-PCR analysis for Inam mRNA induction. Results are presented relative to the expression of Gapdh mRNA. Represented as means ± SD. *p
Figure Legend Snippet: Induction of Inam mRNAs in DCs co-cultured with human cancer cells. Dectin-1-dependent induction of Inam ( Fam26f ) mRNA by a human cancer cell line. Mouse splenic CD11c + cells (3 × 10 5 cells), either from WT mice or Dectin-1 −/− mice, were co-cultured with HBC4 cells (1 × 10 4 cells) for 4 hr and total RNA was subjected to qRT-PCR analysis for Inam mRNA induction. Results are presented relative to the expression of Gapdh mRNA. Represented as means ± SD. *p

Techniques Used: Cell Culture, Mouse Assay, Quantitative RT-PCR, Expressing

Minor effects of Dectin-1 expressed in NK cells on the tumor killing activity. ( A ) In vitro killing activity of purified NK cells from WT or Dectin-1 −/− mice against B16F1 cells. Represented as means ± SD. E/T: effector/target cell ratio. In in vitro killing assays, 1 × 10 4 of 51 Cr-labeled B16F1 cells were used. In vitro killing assays were performed at least three times, and the results were highly reproducible. ( B ) qRT-PCR analysis of Dectin-1 mRNA in WT splenic CD11b + cells, CD11c + cells and NK cells, and MEFs. Results are presented as relative to the expression of Gapdh mRNA. Represented as means ± SD. ND, none detected. ( C ) Expression levels of Dectin-1 mRNA in myeloid cells from liver and lungs. CD11b + F4/80 + cells and CD11c + cells from liver and lung were isolated by cell sorting. Results are presented as relative to the expression of Gapdh mRNA. Represented as means ± SD. DOI: http://dx.doi.org/10.7554/eLife.04177.011
Figure Legend Snippet: Minor effects of Dectin-1 expressed in NK cells on the tumor killing activity. ( A ) In vitro killing activity of purified NK cells from WT or Dectin-1 −/− mice against B16F1 cells. Represented as means ± SD. E/T: effector/target cell ratio. In in vitro killing assays, 1 × 10 4 of 51 Cr-labeled B16F1 cells were used. In vitro killing assays were performed at least three times, and the results were highly reproducible. ( B ) qRT-PCR analysis of Dectin-1 mRNA in WT splenic CD11b + cells, CD11c + cells and NK cells, and MEFs. Results are presented as relative to the expression of Gapdh mRNA. Represented as means ± SD. ND, none detected. ( C ) Expression levels of Dectin-1 mRNA in myeloid cells from liver and lungs. CD11b + F4/80 + cells and CD11c + cells from liver and lung were isolated by cell sorting. Results are presented as relative to the expression of Gapdh mRNA. Represented as means ± SD. DOI: http://dx.doi.org/10.7554/eLife.04177.011

Techniques Used: Activity Assay, In Vitro, Purification, Mouse Assay, Labeling, Quantitative RT-PCR, Expressing, Isolation, FACS

Expression levels of cytotoxic mediators and inflammatory cytokines in co-culture system. NK cells (WT; 1 × 10 5 cells) and WT or Dectin-1 −/− splenic CD11c + cells (3 × 10 5 cells) were co-cultured with B16F1 cells (1 × 10 4 cells). Total RNA was isolated at time zero and 4 hr after the co-culture and then mRNA expression levels for Granzyme B ( Gzmb ), Perforin-1 ( Prf1 ), IFN-γ ( Ifng ), IL-6 ( Il6 ), and TNF-α ( Tnf ) were analyzed by qRT-PCR analysis. Results are presented relative to the expression of Gapdh mRNA. Represented as means ± SD. DOI: http://dx.doi.org/10.7554/eLife.04177.016
Figure Legend Snippet: Expression levels of cytotoxic mediators and inflammatory cytokines in co-culture system. NK cells (WT; 1 × 10 5 cells) and WT or Dectin-1 −/− splenic CD11c + cells (3 × 10 5 cells) were co-cultured with B16F1 cells (1 × 10 4 cells). Total RNA was isolated at time zero and 4 hr after the co-culture and then mRNA expression levels for Granzyme B ( Gzmb ), Perforin-1 ( Prf1 ), IFN-γ ( Ifng ), IL-6 ( Il6 ), and TNF-α ( Tnf ) were analyzed by qRT-PCR analysis. Results are presented relative to the expression of Gapdh mRNA. Represented as means ± SD. DOI: http://dx.doi.org/10.7554/eLife.04177.016

Techniques Used: Expressing, Co-Culture Assay, Cell Culture, Isolation, Quantitative RT-PCR

Induction of Il15 or Il15ra mRNAs in DCs co-cultured with B16F1 cells. Il15 (left panel) or Il15ra (right panel) mRNA were monitored by qRT-PCR analysis as described in Figure 4D . Results are presented relative to the expression of Gapdh mRNA. Represented as means ± SD. *p
Figure Legend Snippet: Induction of Il15 or Il15ra mRNAs in DCs co-cultured with B16F1 cells. Il15 (left panel) or Il15ra (right panel) mRNA were monitored by qRT-PCR analysis as described in Figure 4D . Results are presented relative to the expression of Gapdh mRNA. Represented as means ± SD. *p

Techniques Used: Cell Culture, Quantitative RT-PCR, Expressing

24) Product Images from "Bisphenol A Exposure Enhances Atherosclerosis in WHHL Rabbits"

Article Title: Bisphenol A Exposure Enhances Atherosclerosis in WHHL Rabbits

Journal: PLoS ONE

doi: 10.1371/journal.pone.0110977

ELISA analysis of serum cytokines levels in WHHL rabbits. (A) TNF-α serum concentrations at 0 and 12 weeks. (B) IL-6 serum concentrations at 0 and 12 weeks. Data are expressed as the means ± SD, n = 6 for each group. * p
Figure Legend Snippet: ELISA analysis of serum cytokines levels in WHHL rabbits. (A) TNF-α serum concentrations at 0 and 12 weeks. (B) IL-6 serum concentrations at 0 and 12 weeks. Data are expressed as the means ± SD, n = 6 for each group. * p

Techniques Used: Enzyme-linked Immunosorbent Assay

25) Product Images from "TMPyP4 promotes cancer cell migration at low doses, but induces cell death at high doses"

Article Title: TMPyP4 promotes cancer cell migration at low doses, but induces cell death at high doses

Journal: Scientific Reports

doi: 10.1038/srep26592

The effect of TMPyP4 on cells adhesion to extracellular matrix. ( A–D ) TMPyP4 increase cell adhesion to extracellular matrix. Indicated cell lines were used. ( E ) qRT-PCR showed the abundance of MUC5B mRNA in A549 cancer cell line (NC) and A549 cells transfected with siRNA targeting MUC5B (siRNA_1 and siRNA_2). ( F ) MUC5B knock-down suppress the adhesion of A549 cancer cells to extracellular matrix. MUC5B deficient cells showed no increase of cell adhesion upon TMPyP4 treatment. Values are average ± SD of three independent experiments.
Figure Legend Snippet: The effect of TMPyP4 on cells adhesion to extracellular matrix. ( A–D ) TMPyP4 increase cell adhesion to extracellular matrix. Indicated cell lines were used. ( E ) qRT-PCR showed the abundance of MUC5B mRNA in A549 cancer cell line (NC) and A549 cells transfected with siRNA targeting MUC5B (siRNA_1 and siRNA_2). ( F ) MUC5B knock-down suppress the adhesion of A549 cancer cells to extracellular matrix. MUC5B deficient cells showed no increase of cell adhesion upon TMPyP4 treatment. Values are average ± SD of three independent experiments.

Techniques Used: Quantitative RT-PCR, Transfection

26) Product Images from "Small GTP-binding protein PdRanBP regulates vascular tissue development in poplar"

Article Title: Small GTP-binding protein PdRanBP regulates vascular tissue development in poplar

Journal: BMC Genetics

doi: 10.1186/s12863-016-0403-4

qRT-PCR analysis of the expression of PdRanBP in different vascular tissues and organs of P. deltoides , and detection of immediate and stable expression of GFP -tagged PdRanBP . a , b qRT-PCR analysis of PdRanBP expression in the vascular tissues and other organs of P. deltoides during secondary cell wall development. Aliquots of 1000 ng total RNA were reverse-transcribed into cDNA. The signals were normalized to the constitutively expressed poplar α-tubulin ( TUA1 ) ( a ) and Ubiquitin ( UBQ1 ) ( b ) genes. The values are the mean ± standard error (SE) of three replicates. PdRanBP was predominantly expressed in the leaf buds, immature xylem and immature phloem of P. deltoides . c Nuclear localization of EGFP-PdRanBP fusion protein in onion epidermal cells. Dark-field images were captured for green fluorescence ( e and f ), GFP-only control (g) and the corresponding bright-field images for e , f, g are a, b, c . Bright-field images ( h ) were captured for cell morphology, and the corresponding dark-field images for h is d . i and j , Nuclei counterstained with 4′, 6-diamidino-2-phenylindole (DAPI); the corresponding GFP-only control images of i and j are shown in k . The scale bars are 200 μm in a , c, d, e, g, h, i and k , and 800 μm in b, f , and j
Figure Legend Snippet: qRT-PCR analysis of the expression of PdRanBP in different vascular tissues and organs of P. deltoides , and detection of immediate and stable expression of GFP -tagged PdRanBP . a , b qRT-PCR analysis of PdRanBP expression in the vascular tissues and other organs of P. deltoides during secondary cell wall development. Aliquots of 1000 ng total RNA were reverse-transcribed into cDNA. The signals were normalized to the constitutively expressed poplar α-tubulin ( TUA1 ) ( a ) and Ubiquitin ( UBQ1 ) ( b ) genes. The values are the mean ± standard error (SE) of three replicates. PdRanBP was predominantly expressed in the leaf buds, immature xylem and immature phloem of P. deltoides . c Nuclear localization of EGFP-PdRanBP fusion protein in onion epidermal cells. Dark-field images were captured for green fluorescence ( e and f ), GFP-only control (g) and the corresponding bright-field images for e , f, g are a, b, c . Bright-field images ( h ) were captured for cell morphology, and the corresponding dark-field images for h is d . i and j , Nuclei counterstained with 4′, 6-diamidino-2-phenylindole (DAPI); the corresponding GFP-only control images of i and j are shown in k . The scale bars are 200 μm in a , c, d, e, g, h, i and k , and 800 μm in b, f , and j

Techniques Used: Quantitative RT-PCR, Expressing, Fluorescence

27) Product Images from "Overexpression of the Eggplant (Solanum melongena) NAC Family Transcription Factor SmNAC Suppresses Resistance to Bacterial Wilt"

Article Title: Overexpression of the Eggplant (Solanum melongena) NAC Family Transcription Factor SmNAC Suppresses Resistance to Bacterial Wilt

Journal: Scientific Reports

doi: 10.1038/srep31568

qRT-PCR analysis of the expression of defense signaling genes in transgenic eggplants. Quantitative reverse-transcription PCR (qPCR) was performed using a SYBR Premix Ex Taq kit (TaKaRa, Dalian, China), following the manufacturer’s protocols. Triplicate qPCR reactions were performed for each sample and the relative gene expression data was analyzed using the 2 −ΔΔ Ct method. ( a ) qRT-PCR analysis of defense signaling genes in SmNAC oveexpressing plants. CK represents non-transgenic plants from the E-31 line, whereas 1–3 show the SmNAC overexpressing transgenic T 0 plants EGT 0–87 , EGT 0–145 , and EGT 0–204 , and 4–6 show the SmNAC overexpressing transgenic T 1 plants EGT 1–87 , EGT 1–145 , and EGT 1–204 . ( b ) qRT-PCR analysis of defense signaling genes in RNAi- SmNAC plants. CK represents non-transgenic plants from line E-32. 1–5 show the RNAi- SmNAC transgenic plants (T0) RNAi-1, RNAi-2, RNAi-3, RNAi-4, and RNAi-5.
Figure Legend Snippet: qRT-PCR analysis of the expression of defense signaling genes in transgenic eggplants. Quantitative reverse-transcription PCR (qPCR) was performed using a SYBR Premix Ex Taq kit (TaKaRa, Dalian, China), following the manufacturer’s protocols. Triplicate qPCR reactions were performed for each sample and the relative gene expression data was analyzed using the 2 −ΔΔ Ct method. ( a ) qRT-PCR analysis of defense signaling genes in SmNAC oveexpressing plants. CK represents non-transgenic plants from the E-31 line, whereas 1–3 show the SmNAC overexpressing transgenic T 0 plants EGT 0–87 , EGT 0–145 , and EGT 0–204 , and 4–6 show the SmNAC overexpressing transgenic T 1 plants EGT 1–87 , EGT 1–145 , and EGT 1–204 . ( b ) qRT-PCR analysis of defense signaling genes in RNAi- SmNAC plants. CK represents non-transgenic plants from line E-32. 1–5 show the RNAi- SmNAC transgenic plants (T0) RNAi-1, RNAi-2, RNAi-3, RNAi-4, and RNAi-5.

Techniques Used: Quantitative RT-PCR, Expressing, Transgenic Assay, Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

28) Product Images from "A Malus Crabapple Chalcone Synthase Gene, McCHS, Regulates Red Petal Color and Flavonoid Biosynthesis"

Article Title: A Malus Crabapple Chalcone Synthase Gene, McCHS, Regulates Red Petal Color and Flavonoid Biosynthesis

Journal: PLoS ONE

doi: 10.1371/journal.pone.0110570

Relative expression profile of downstream genes of McCHS during flower developmental stages. Real-time PCR was used to analyze the expression patterns of McCHS , McF3H , McF3′H , McDFR , McANS and McUFGT in petals of Malus cultivars ‘Royalty’ and ‘Radiant’, ‘Flame’. All real time-PCR reactions were normalized using the Ct value corresponding to a Malus crabapple 18S ribosomal RNA gene (DQ341382). Stages referred to on the x axis are: (I) 6 days before full bloom; (II) 3 days before full bloom; (III) 1 day before full bloom; (IV) full bloom; and (V) 3 days after full bloom. Error bars correspond to the standard error of the mean ± SE of three replicate reactions.
Figure Legend Snippet: Relative expression profile of downstream genes of McCHS during flower developmental stages. Real-time PCR was used to analyze the expression patterns of McCHS , McF3H , McF3′H , McDFR , McANS and McUFGT in petals of Malus cultivars ‘Royalty’ and ‘Radiant’, ‘Flame’. All real time-PCR reactions were normalized using the Ct value corresponding to a Malus crabapple 18S ribosomal RNA gene (DQ341382). Stages referred to on the x axis are: (I) 6 days before full bloom; (II) 3 days before full bloom; (III) 1 day before full bloom; (IV) full bloom; and (V) 3 days after full bloom. Error bars correspond to the standard error of the mean ± SE of three replicate reactions.

Techniques Used: Expressing, Real-time Polymerase Chain Reaction

29) Product Images from "The novel circCLK3/miR-320a/FoxM1 axis promotes cervical cancer progression"

Article Title: The novel circCLK3/miR-320a/FoxM1 axis promotes cervical cancer progression

Journal: Cell Death & Disease

doi: 10.1038/s41419-019-2183-z

CircCLK3 expression is relatively highly expressed in cervical cancer tissues and predicts poor prognosis. a CircRNA sequencing was adapted to detect the differentially expressed circRNAs between cervical cancer tissues and matched normal tissues. b The genomic loci of the CLK3 gene and circCLK3. Sanger sequencing confirmed the head-to-hail splicing. c The expression level of circCLK3 in 5 cervical cancer cell lines relative to HeLa cell. d The gel electrophoresis validated the existence of circCLK3. Divergent primers amplified circCLK3 in cDNA but not gDNA. e qRT-PCR results of U6, GAPDH, and circCLK3 expressions in cell nuclei and cytoplasm in SiHa and HeLa cells. f RNA-FISH assays were performed to identify the subcellular location of circCLK3 with the cy3-labeled circCLK3 probe. The results showed that most of circCLK3 was located in cytoplasm in SiHa and HeLa cells. g qRT-PCR results of circCLK3 and CLK3 expression after treatment with actinomycin D at the indicated time points in SiHa and HeLa cells. h qRT-PCR results of circCLK3 and CLK3 expression after treatment with RNase R in SiHa and HeLa cells. i CircCLK3 expression was significantly higher in 48 paired fresh frozen cervical cancer tissues compared with adjacent normal tissues. j , k Kaplan–Meier Plotter analysis of the correlation between circCLK3 level with overall survive or disease-free survive of cervical cancer patients. Data are reported as means ± standard deviation of three independent experiments. * p
Figure Legend Snippet: CircCLK3 expression is relatively highly expressed in cervical cancer tissues and predicts poor prognosis. a CircRNA sequencing was adapted to detect the differentially expressed circRNAs between cervical cancer tissues and matched normal tissues. b The genomic loci of the CLK3 gene and circCLK3. Sanger sequencing confirmed the head-to-hail splicing. c The expression level of circCLK3 in 5 cervical cancer cell lines relative to HeLa cell. d The gel electrophoresis validated the existence of circCLK3. Divergent primers amplified circCLK3 in cDNA but not gDNA. e qRT-PCR results of U6, GAPDH, and circCLK3 expressions in cell nuclei and cytoplasm in SiHa and HeLa cells. f RNA-FISH assays were performed to identify the subcellular location of circCLK3 with the cy3-labeled circCLK3 probe. The results showed that most of circCLK3 was located in cytoplasm in SiHa and HeLa cells. g qRT-PCR results of circCLK3 and CLK3 expression after treatment with actinomycin D at the indicated time points in SiHa and HeLa cells. h qRT-PCR results of circCLK3 and CLK3 expression after treatment with RNase R in SiHa and HeLa cells. i CircCLK3 expression was significantly higher in 48 paired fresh frozen cervical cancer tissues compared with adjacent normal tissues. j , k Kaplan–Meier Plotter analysis of the correlation between circCLK3 level with overall survive or disease-free survive of cervical cancer patients. Data are reported as means ± standard deviation of three independent experiments. * p

Techniques Used: Expressing, Sequencing, Nucleic Acid Electrophoresis, Amplification, Quantitative RT-PCR, Fluorescence In Situ Hybridization, Labeling, Standard Deviation

30) Product Images from "Synergistic anti-proliferative effects of gambogic acid with docetaxel in gastrointestinal cancer cell lines"

Article Title: Synergistic anti-proliferative effects of gambogic acid with docetaxel in gastrointestinal cancer cell lines

Journal: BMC Complementary and Alternative Medicine

doi: 10.1186/1472-6882-12-58

GA down-regulates the expression of genes involved in Doc sensitivity . The mRNA expression of genes involved in Doc sensitivity, including β-tubulin III, tau and survivin, in BGC-823 cells was determined by qRT-PCR after 48 h of GA (0.25 μM) treatment. The mRNA expression of target genes was normalized to a control. * P
Figure Legend Snippet: GA down-regulates the expression of genes involved in Doc sensitivity . The mRNA expression of genes involved in Doc sensitivity, including β-tubulin III, tau and survivin, in BGC-823 cells was determined by qRT-PCR after 48 h of GA (0.25 μM) treatment. The mRNA expression of target genes was normalized to a control. * P

Techniques Used: Expressing, Quantitative RT-PCR

31) Product Images from "Functional analysis of the GmESR1 gene associated with soybean regeneration"

Article Title: Functional analysis of the GmESR1 gene associated with soybean regeneration

Journal: PLoS ONE

doi: 10.1371/journal.pone.0175656

Expression patterns of GmESR1 in different organs of the soybean cultivar ‘Dongnong 50’. Transcript abundances were normalized against the reference gene GmActin4 . Bars and error bars represent the mean ± standard error of three experiments with independent RNA extractions.
Figure Legend Snippet: Expression patterns of GmESR1 in different organs of the soybean cultivar ‘Dongnong 50’. Transcript abundances were normalized against the reference gene GmActin4 . Bars and error bars represent the mean ± standard error of three experiments with independent RNA extractions.

Techniques Used: Expressing

Analysis of the purified recombinant GmESR1 protein. (A) The recombinant GmESR1 protein, induced with 0.5 mM IPTG at 37°C for 2, 4, 6, and 8 h in E . coli BL21 competent cells. (B) SDS-PAGE analysis of the purified recombinant GmESR1 protein using the His-Bind kit.
Figure Legend Snippet: Analysis of the purified recombinant GmESR1 protein. (A) The recombinant GmESR1 protein, induced with 0.5 mM IPTG at 37°C for 2, 4, 6, and 8 h in E . coli BL21 competent cells. (B) SDS-PAGE analysis of the purified recombinant GmESR1 protein using the His-Bind kit.

Techniques Used: Purification, Recombinant, SDS Page

Nucleotide and amino acid sequences of GmESR1 . Putative phosphorylation sites are marked in bold italics. The YRG element and RAYD element are highlighted by shading. The α-helix and β-sheets are underlined. Amino acid and base pair numbers are shown on the left.
Figure Legend Snippet: Nucleotide and amino acid sequences of GmESR1 . Putative phosphorylation sites are marked in bold italics. The YRG element and RAYD element are highlighted by shading. The α-helix and β-sheets are underlined. Amino acid and base pair numbers are shown on the left.

Techniques Used:

Sequence-specific binding of GmESR1 to the GCC-box element. (A) Nucleotide sequences of the GCC-box and mGCC-box probes. (B) Electrophoretic mobility shift assay (EMSA) showed sequence-specific binding to the GCC-box of the recombinant GmESR1 protein. Lane 1, EMSA performed with only the free GCC probes; lane 2, labeled GCC probe and GmESR1 protein; lane 3, titration with a cold GCC sequence as a competitor; lane 4, titration with a cold mGCC-box sequence as a competitor; lane 5, labeled mGCC probe and GmESR1 protein.
Figure Legend Snippet: Sequence-specific binding of GmESR1 to the GCC-box element. (A) Nucleotide sequences of the GCC-box and mGCC-box probes. (B) Electrophoretic mobility shift assay (EMSA) showed sequence-specific binding to the GCC-box of the recombinant GmESR1 protein. Lane 1, EMSA performed with only the free GCC probes; lane 2, labeled GCC probe and GmESR1 protein; lane 3, titration with a cold GCC sequence as a competitor; lane 4, titration with a cold mGCC-box sequence as a competitor; lane 5, labeled mGCC probe and GmESR1 protein.

Techniques Used: Sequencing, Binding Assay, Electrophoretic Mobility Shift Assay, Recombinant, Labeling, Titration

Identification and analysis of mutant atesr1 and GmESR1 -ox Arabidopsis plants. (A) Phenotypes of homozygous atesr1 mutant (HM), heterozygous atesr1 mutant (HZ) and wild-type plant (WT). Scale bar = 1.0 cm. (B) PCR result s for the genotyping assay to identify atesr1 mutant plants. (C) Identification of transgenic GmESR1 -ox Arabidopsis plants using qRT-PCR. Transcript abundances were normalized against the reference gene AtActin8 . (D) Germination of mutant atesr1 , WT, and GmESR1 -ox Arabidopsis seeds. Two independent GmESR1 -ox lines are included. (E) Germination of atesr1 , WT, and GmESR1 -ox seeds after 2.5 and 6.5 d. (F) Comparison of GmESR1 -ox, WT, and atesr1 elongation rates on MS medium 2.5 d after planting. (G) Root length in GmESR1 -ox, WT, and mutant atesr1 plants 2.5 d after planting. (H) Phenotypes of WT, GmESR1 -ox, and atesr1 plants 30 d after transplanting. The experiment was performed on three biological replicates with their respective three technical replicates and statistically analyzed using Student’s t -test (* P
Figure Legend Snippet: Identification and analysis of mutant atesr1 and GmESR1 -ox Arabidopsis plants. (A) Phenotypes of homozygous atesr1 mutant (HM), heterozygous atesr1 mutant (HZ) and wild-type plant (WT). Scale bar = 1.0 cm. (B) PCR result s for the genotyping assay to identify atesr1 mutant plants. (C) Identification of transgenic GmESR1 -ox Arabidopsis plants using qRT-PCR. Transcript abundances were normalized against the reference gene AtActin8 . (D) Germination of mutant atesr1 , WT, and GmESR1 -ox Arabidopsis seeds. Two independent GmESR1 -ox lines are included. (E) Germination of atesr1 , WT, and GmESR1 -ox seeds after 2.5 and 6.5 d. (F) Comparison of GmESR1 -ox, WT, and atesr1 elongation rates on MS medium 2.5 d after planting. (G) Root length in GmESR1 -ox, WT, and mutant atesr1 plants 2.5 d after planting. (H) Phenotypes of WT, GmESR1 -ox, and atesr1 plants 30 d after transplanting. The experiment was performed on three biological replicates with their respective three technical replicates and statistically analyzed using Student’s t -test (* P

Techniques Used: Mutagenesis, Polymerase Chain Reaction, Genotyping Assay, Transgenic Assay, Quantitative RT-PCR, Mass Spectrometry

Characterization of GmESR1 . (A) Phylogenetic analysis of GmESR1 with 20 other ESR1 proteins The GenBank accession numbers are as follows: Glycine max [JN590243], Vigna angularis [KOM39777], Cicer arietinum [XP_004489775], Medicago truncatula [XP_003613106], Vitis vinifera [XP_002271778], Theobroma cacao [XP_007044810], Populus angustifolia [AGA18050], Populus trichocarpa [XP_002314597], Solanum tuberosum [XP_006357626], Cucumis melo [XM_008446150], Amborella trichopoda [XM_006852288], Arabidopsis thaliana [NM_101169], Brassica oleracea [XM_013746383], Brassica rapa [XM_009119902], Oryza sativa [Oryza sativa Japonica Group] [NP_001047305], Sorghum bicolor [XM_002452333], Zea mays [NM_001153873], Capsella rubella [EOA21975], Cucumis sativus [XP_004152327] and Prunus persica [EMJ26264]. (B) Alignment of amino acid sequences of GmESR1 and the four most similar ESR1 proteins. The YRG element and RAYD element are indicated by horizontal lines above the sequence. Amino acid numbers are indicated on the right. (C) The predicted three-dimensional structure of GmESR1. (D) The conserved domain of the GmESR1 protein. The predicted GmESR1 protein contains a conserved domain at amino acids 51–107 that belongs to the AP2 superfamily.
Figure Legend Snippet: Characterization of GmESR1 . (A) Phylogenetic analysis of GmESR1 with 20 other ESR1 proteins The GenBank accession numbers are as follows: Glycine max [JN590243], Vigna angularis [KOM39777], Cicer arietinum [XP_004489775], Medicago truncatula [XP_003613106], Vitis vinifera [XP_002271778], Theobroma cacao [XP_007044810], Populus angustifolia [AGA18050], Populus trichocarpa [XP_002314597], Solanum tuberosum [XP_006357626], Cucumis melo [XM_008446150], Amborella trichopoda [XM_006852288], Arabidopsis thaliana [NM_101169], Brassica oleracea [XM_013746383], Brassica rapa [XM_009119902], Oryza sativa [Oryza sativa Japonica Group] [NP_001047305], Sorghum bicolor [XM_002452333], Zea mays [NM_001153873], Capsella rubella [EOA21975], Cucumis sativus [XP_004152327] and Prunus persica [EMJ26264]. (B) Alignment of amino acid sequences of GmESR1 and the four most similar ESR1 proteins. The YRG element and RAYD element are indicated by horizontal lines above the sequence. Amino acid numbers are indicated on the right. (C) The predicted three-dimensional structure of GmESR1. (D) The conserved domain of the GmESR1 protein. The predicted GmESR1 protein contains a conserved domain at amino acids 51–107 that belongs to the AP2 superfamily.

Techniques Used: Sequencing

Analysis of GmESR1 transgenic soybean plants. (A) Relative expression level of GmESR1 in control group and four independent GmESR1 -ox lines. Transcript abundance was normalized against the reference gene GmActin4 . (B) Comparison of elongation rate in GmESR1 -ox and control soybean plants during germination 5 d after planting. (C) Comparison of shoot elongation rate in GmESR1 -ox and control soybean seedlings 15 d after planting. (D) Comparison of root elongation in GmESR1 -ox and control soybean seedlings 15 d after planting. The experiments were performed on three biological replicates with their respective three technical replicates. Error bars represent the standard error of the mean. Scale bars = 1.0 cm. (E) Comparison of the bud cells in GmESR1 -ox and control soybean plants during the bud induction. Scale bars = 5μm.
Figure Legend Snippet: Analysis of GmESR1 transgenic soybean plants. (A) Relative expression level of GmESR1 in control group and four independent GmESR1 -ox lines. Transcript abundance was normalized against the reference gene GmActin4 . (B) Comparison of elongation rate in GmESR1 -ox and control soybean plants during germination 5 d after planting. (C) Comparison of shoot elongation rate in GmESR1 -ox and control soybean seedlings 15 d after planting. (D) Comparison of root elongation in GmESR1 -ox and control soybean seedlings 15 d after planting. The experiments were performed on three biological replicates with their respective three technical replicates. Error bars represent the standard error of the mean. Scale bars = 1.0 cm. (E) Comparison of the bud cells in GmESR1 -ox and control soybean plants during the bud induction. Scale bars = 5μm.

Techniques Used: Transgenic Assay, Expressing

32) Product Images from "CircPSMC3 suppresses the proliferation and metastasis of gastric cancer by acting as a competitive endogenous RNA through sponging miR-296-5p"

Article Title: CircPSMC3 suppresses the proliferation and metastasis of gastric cancer by acting as a competitive endogenous RNA through sponging miR-296-5p

Journal: Molecular Cancer

doi: 10.1186/s12943-019-0958-6

CircPSMC3 expression is down regulated in clinical GC specimens and cell lines. a Scatter plot analyses of circRNAs microarray data showing differentially expressed circRNA in lymph node metastasis group and normal group . High expression level is indicated by “red” and low levels by “green”. b Scatter plot analyses of circRNA microarray data showing differentially expressed circRNAs in lymph node metastasis group and no lymph node metastasis group. c GO analysis of circRNAs in lymph node metastasis group and normal group. d GO analysis of circRNAs in lymph node metastasis group and no lymph node metastasis group. e The spliced mature sequence length of circPSMC3 derived from the PSMC3 gene is 502 bp. f QRT-PCR for the abundance of circPSMC3 in GC cells treated with RNase R. g QRT-PCR for the abundance of PSMC3 mRNA in GC cells treated with RNase R. h CircPSMC3 expressions were evaluated using qRT-PCR in 106 pairs of gastric cancer plasmas and 21 pairs of normal plasmas. i CircPSMC3 expressions were measured using qRT-PCR in 106 pairs of gastric cancer (Tumor) and matched noncancerous tissue (Normal). j The expressions of circPSMC3 were evaluated in cell lines using qRT-PCR. k The area under the ROC curve (AUC) in distinguishing GC plasmas and normal ones was 0.9326. l Kaplan Meier survival curve showed the relationship between circPSMC3 and survival rates. ** p
Figure Legend Snippet: CircPSMC3 expression is down regulated in clinical GC specimens and cell lines. a Scatter plot analyses of circRNAs microarray data showing differentially expressed circRNA in lymph node metastasis group and normal group . High expression level is indicated by “red” and low levels by “green”. b Scatter plot analyses of circRNA microarray data showing differentially expressed circRNAs in lymph node metastasis group and no lymph node metastasis group. c GO analysis of circRNAs in lymph node metastasis group and normal group. d GO analysis of circRNAs in lymph node metastasis group and no lymph node metastasis group. e The spliced mature sequence length of circPSMC3 derived from the PSMC3 gene is 502 bp. f QRT-PCR for the abundance of circPSMC3 in GC cells treated with RNase R. g QRT-PCR for the abundance of PSMC3 mRNA in GC cells treated with RNase R. h CircPSMC3 expressions were evaluated using qRT-PCR in 106 pairs of gastric cancer plasmas and 21 pairs of normal plasmas. i CircPSMC3 expressions were measured using qRT-PCR in 106 pairs of gastric cancer (Tumor) and matched noncancerous tissue (Normal). j The expressions of circPSMC3 were evaluated in cell lines using qRT-PCR. k The area under the ROC curve (AUC) in distinguishing GC plasmas and normal ones was 0.9326. l Kaplan Meier survival curve showed the relationship between circPSMC3 and survival rates. ** p

Techniques Used: Expressing, Microarray, Sequencing, Derivative Assay, Quantitative RT-PCR

33) Product Images from "Stromal interaction molecule 1 regulates growth, cell cycle, and apoptosis of human tongue squamous carcinoma cells"

Article Title: Stromal interaction molecule 1 regulates growth, cell cycle, and apoptosis of human tongue squamous carcinoma cells

Journal: Bioscience Reports

doi: 10.1042/BSR20160519

STIM1 knockdown induces apoptosis in Tca-8113 cells ( A ) Representative images showing STIM1 knockdown induces apoptosis in Tca-8113 cells. Tca-8113 cells were infected shCtrl or shSTIM1 for 48 h and then cell apoptosis was determined by Annexin V staining and flow cytometry. R3 region indicates the percentage of apoptotic cells. ( B ) Quantitation of cell apoptosis induced by knockdown of STIM1 in (A); *** P
Figure Legend Snippet: STIM1 knockdown induces apoptosis in Tca-8113 cells ( A ) Representative images showing STIM1 knockdown induces apoptosis in Tca-8113 cells. Tca-8113 cells were infected shCtrl or shSTIM1 for 48 h and then cell apoptosis was determined by Annexin V staining and flow cytometry. R3 region indicates the percentage of apoptotic cells. ( B ) Quantitation of cell apoptosis induced by knockdown of STIM1 in (A); *** P

Techniques Used: Infection, Staining, Flow Cytometry, Cytometry, Quantitation Assay

STIM1 knockdown inhibits Tca-8113 cell proliferation ( A ) Representative images of Tca-8113 cells infected with shCtrl (top) and shSTIM1 (bottom) via multiparametric HCS every day for 5 days. ( B – C ) STIM1 knockdown represses cell proliferation of Tca-8113 cells. Tca-8113 cells infected with shCtrl and shSTIM1, and cell number was analyzed everyday by HCS.
Figure Legend Snippet: STIM1 knockdown inhibits Tca-8113 cell proliferation ( A ) Representative images of Tca-8113 cells infected with shCtrl (top) and shSTIM1 (bottom) via multiparametric HCS every day for 5 days. ( B – C ) STIM1 knockdown represses cell proliferation of Tca-8113 cells. Tca-8113 cells infected with shCtrl and shSTIM1, and cell number was analyzed everyday by HCS.

Techniques Used: Infection

Clonogenic ability was suppressed in Tca-8113 cells by STIM1 knockdown ( A ) Pictures of colony formation by Tca-8113 cells infected with lentivirus expression either shCtrl or shSTIM1 using Giemsa staining on the 14th day. ( B ) Quantitation of clone number formed by Tca-8113 cells infected with lentivirus expression either shCtrl or shSTIM1. The results represent mean ± S.E.M.; *** P
Figure Legend Snippet: Clonogenic ability was suppressed in Tca-8113 cells by STIM1 knockdown ( A ) Pictures of colony formation by Tca-8113 cells infected with lentivirus expression either shCtrl or shSTIM1 using Giemsa staining on the 14th day. ( B ) Quantitation of clone number formed by Tca-8113 cells infected with lentivirus expression either shCtrl or shSTIM1. The results represent mean ± S.E.M.; *** P

Techniques Used: Infection, Expressing, Staining, Quantitation Assay

Knockdown of STIM1 regulates cell cycle of Tca-8113 cells ( A ) Representative result showing STIM1 knockdown represses cell cycle in Tca-8113 cells. Tca-8113 cells were infected with shCtrl or shSTIM1 lentivirus for 48 h and then flow cytometry was performed to analyze cell cycle. ( B ) STIM1 knockdown induces cell cycle arrest at G 1 phase. Tca-8113 cells were infected with shCtrl or shSTIM1 lentivirus for 48 h and then flow cytometry was performed to analyze cell cycle; *** P
Figure Legend Snippet: Knockdown of STIM1 regulates cell cycle of Tca-8113 cells ( A ) Representative result showing STIM1 knockdown represses cell cycle in Tca-8113 cells. Tca-8113 cells were infected with shCtrl or shSTIM1 lentivirus for 48 h and then flow cytometry was performed to analyze cell cycle. ( B ) STIM1 knockdown induces cell cycle arrest at G 1 phase. Tca-8113 cells were infected with shCtrl or shSTIM1 lentivirus for 48 h and then flow cytometry was performed to analyze cell cycle; *** P

Techniques Used: Infection, Flow Cytometry, Cytometry

Knockdown of STIM1 disruption multiple critical pathways in human tongue squamous carcinoma cells ( A ) Heatmap representation of 1841 genes (833 genes up-regulated and 1008 genes down-regulated) with differential expressions in Tca-8113 cells infected with lentivirus expressing either shCtrl or shSTIM1 (criteria: P
Figure Legend Snippet: Knockdown of STIM1 disruption multiple critical pathways in human tongue squamous carcinoma cells ( A ) Heatmap representation of 1841 genes (833 genes up-regulated and 1008 genes down-regulated) with differential expressions in Tca-8113 cells infected with lentivirus expressing either shCtrl or shSTIM1 (criteria: P

Techniques Used: Infection, Expressing

STIM1 was expressed and knocked down in human tongue squamous carcinoma cells ( A ) Expression of STIM1 mRNA was measured by qRT-PCR in two tongue squamous carcinoma cell lines, Tca-8113 and CAL27. The relative expression of STIM1 is shown as Δ C T ( C T STIM1 − C T GAPDH ). ( B ) Representative image showing lentivirus infection in Tca-8113 cells. Tca-8113 cells were infected with shSTIM1 and shCtrl lentivirus for 48 h, and then the efficiency of infection was assessed by light microscopy and fluorescent microscopy at the third day infection. ( C ) Knockdown of STIM1 in Tca-8113 cells. Tca-8113 cells were infected with shSTIM1 and shCtrl lentivirus for 48 h, and then qRT-PCR analysis was performed to analyze the mRNA level of STIM1 in Tca-8113 cells; *** P
Figure Legend Snippet: STIM1 was expressed and knocked down in human tongue squamous carcinoma cells ( A ) Expression of STIM1 mRNA was measured by qRT-PCR in two tongue squamous carcinoma cell lines, Tca-8113 and CAL27. The relative expression of STIM1 is shown as Δ C T ( C T STIM1 − C T GAPDH ). ( B ) Representative image showing lentivirus infection in Tca-8113 cells. Tca-8113 cells were infected with shSTIM1 and shCtrl lentivirus for 48 h, and then the efficiency of infection was assessed by light microscopy and fluorescent microscopy at the third day infection. ( C ) Knockdown of STIM1 in Tca-8113 cells. Tca-8113 cells were infected with shSTIM1 and shCtrl lentivirus for 48 h, and then qRT-PCR analysis was performed to analyze the mRNA level of STIM1 in Tca-8113 cells; *** P

Techniques Used: Expressing, Quantitative RT-PCR, Infection, Light Microscopy, Microscopy

34) Product Images from "De novo Transcriptome Assembly of Common Wild Rice (Oryza rufipogon Griff.) and Discovery of Drought-Response Genes in Root Tissue Based on Transcriptomic Data"

Article Title: De novo Transcriptome Assembly of Common Wild Rice (Oryza rufipogon Griff.) and Discovery of Drought-Response Genes in Root Tissue Based on Transcriptomic Data

Journal: PLoS ONE

doi: 10.1371/journal.pone.0131455

Characteristics of the BLAST matches of the transcriptome unigenes. (A) E-value and (B) Similarity distribution of the top BLAST hits for each unigene in the NR database, with a cut-off E-value of 1.0E-5. (C) Species distribution of the BLAST hits for each unigene in the NT database.
Figure Legend Snippet: Characteristics of the BLAST matches of the transcriptome unigenes. (A) E-value and (B) Similarity distribution of the top BLAST hits for each unigene in the NR database, with a cut-off E-value of 1.0E-5. (C) Species distribution of the BLAST hits for each unigene in the NT database.

Techniques Used:

Expression characters of four selected unigenes in the roots of PEG-treated plants at 0 h, 24 h, 48 h and 72 h. The relative expression value was calculated using the 2 -ΔΔCt method and 0 h as a control sample.
Figure Legend Snippet: Expression characters of four selected unigenes in the roots of PEG-treated plants at 0 h, 24 h, 48 h and 72 h. The relative expression value was calculated using the 2 -ΔΔCt method and 0 h as a control sample.

Techniques Used: Expressing

Functional classification of the assembled unigenes. (A) Comparison of Gene Ontology (GO) classifications of common wild rice. Unigene numbers assigned to the same GO terms are indicated above the bars; the x-axis indicates the subcategories, and the y-axis indicates the number of genes in a category. (B) Histogram of the Eukaryotic Orthologous Groups (KOG) classification.
Figure Legend Snippet: Functional classification of the assembled unigenes. (A) Comparison of Gene Ontology (GO) classifications of common wild rice. Unigene numbers assigned to the same GO terms are indicated above the bars; the x-axis indicates the subcategories, and the y-axis indicates the number of genes in a category. (B) Histogram of the Eukaryotic Orthologous Groups (KOG) classification.

Techniques Used: Functional Assay

35) Product Images from "Circular RNA hsa_circ_0000885 Levels are Increased in Tissue and Serum Samples from Patients with Osteosarcoma"

Article Title: Circular RNA hsa_circ_0000885 Levels are Increased in Tissue and Serum Samples from Patients with Osteosarcoma

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

doi: 10.12659/MSM.914899

Serum hsa_circ_0000885 levels were increased in serum from patients with osteosarcoma. ( A ) Expression levels of hsa_circ_0000885 in 30 patients with osteosarcoma, 27 patients with benign bone tumors, and 25 age-matched and sex-matched healthy controls were analyzed by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and normalized to the GAPDH level. ( B ) Relative expression of hsa_circ_0000885 in 11 patients with osteosarcoma, Enneking stage I or IIA and 19 patients with osteosarcoma, Enneking stage IB or III. ( C ) Relative expression of hsa_circ_0000885 in 20 patients with osteosarcoma without metastasis and 10 patients with osteosarcoma with lung metastasis. ( D ) Relative expression of hsa_circ_0000885 in 30 patients with osteosarcoma before and after chemotherapy. ( E ) Relative expression of hsa_circ_0000885 in 30 patients with osteosarcoma before and after surgery. ( F ) Receiver operating characteristic (ROC) curve shows the diagnostic value of hsa_circ_0000885 in distinguishing between patients with osteosarcoma from healthy individuals. ( G ) ROC curve shows the diagnostic value of hsa_circ_0000885 in distinguishing between patients with osteosarcoma and benign bone tumors.
Figure Legend Snippet: Serum hsa_circ_0000885 levels were increased in serum from patients with osteosarcoma. ( A ) Expression levels of hsa_circ_0000885 in 30 patients with osteosarcoma, 27 patients with benign bone tumors, and 25 age-matched and sex-matched healthy controls were analyzed by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and normalized to the GAPDH level. ( B ) Relative expression of hsa_circ_0000885 in 11 patients with osteosarcoma, Enneking stage I or IIA and 19 patients with osteosarcoma, Enneking stage IB or III. ( C ) Relative expression of hsa_circ_0000885 in 20 patients with osteosarcoma without metastasis and 10 patients with osteosarcoma with lung metastasis. ( D ) Relative expression of hsa_circ_0000885 in 30 patients with osteosarcoma before and after chemotherapy. ( E ) Relative expression of hsa_circ_0000885 in 30 patients with osteosarcoma before and after surgery. ( F ) Receiver operating characteristic (ROC) curve shows the diagnostic value of hsa_circ_0000885 in distinguishing between patients with osteosarcoma from healthy individuals. ( G ) ROC curve shows the diagnostic value of hsa_circ_0000885 in distinguishing between patients with osteosarcoma and benign bone tumors.

Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR, Diagnostic Assay

Increased hsa_circ_0000885 expression was correlated with poor clinical prognosis. ( A ) Disease-free survival (DFS) in patients with high hsa_circ_0000885 expression and low hsa_circ_0000885 expression (p
Figure Legend Snippet: Increased hsa_circ_0000885 expression was correlated with poor clinical prognosis. ( A ) Disease-free survival (DFS) in patients with high hsa_circ_0000885 expression and low hsa_circ_0000885 expression (p

Techniques Used: Expressing

hsa_circ_0000885 expression was upregulated in osteosarcoma tissue. ( A ) The heatmap shows the circRNA expression profiles of osteosarcoma tissues and adjacent non-tumor tissues. Red and green indicate high and low expression, respectively. ( B ) Expression levels of hsa_circ_0000885 in 50 pairs of osteosarcoma tissues and adjacent non-tumor tissues, analyzed by quantitative real-time polymerase chain reaction (qRT-PCR) and normalized to GAPDH. The results were analyzed using the 2 −ΔΔCT method and compared using paired Student’s t-tests. ( C ) Relative hsa_circ_0000885 expression ratios in osteosarcoma tissues versus adjacent non-tumor tissues shown on the logarithmic scale. Data are shown as the means ± standard deviation (SD). *** P
Figure Legend Snippet: hsa_circ_0000885 expression was upregulated in osteosarcoma tissue. ( A ) The heatmap shows the circRNA expression profiles of osteosarcoma tissues and adjacent non-tumor tissues. Red and green indicate high and low expression, respectively. ( B ) Expression levels of hsa_circ_0000885 in 50 pairs of osteosarcoma tissues and adjacent non-tumor tissues, analyzed by quantitative real-time polymerase chain reaction (qRT-PCR) and normalized to GAPDH. The results were analyzed using the 2 −ΔΔCT method and compared using paired Student’s t-tests. ( C ) Relative hsa_circ_0000885 expression ratios in osteosarcoma tissues versus adjacent non-tumor tissues shown on the logarithmic scale. Data are shown as the means ± standard deviation (SD). *** P

Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Standard Deviation

36) Product Images from "Protective Effects of Sinomenine on CFA-Induced Inflammatory Pain in Rats"

Article Title: Protective Effects of Sinomenine on CFA-Induced Inflammatory Pain in Rats

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

doi: 10.12659/MSM.906726

SIN inhibits COX-2 expression in IP rats. ( A ) The protein level of COX-2 was detected by Western blot analysis. ( B ) The mRNA level of COX-2 was detected by qRT-PCR. Saline: rats were injected with 100 μl saline only; Saline+SIN: rats were injected with 30 mg/kg SIN and 100 μl saline; CFA: rats were injected with 100 μl CFA only; CFA+SIN: rats were injected with 30 mg/kg SIN and 100 μl CFA. * p
Figure Legend Snippet: SIN inhibits COX-2 expression in IP rats. ( A ) The protein level of COX-2 was detected by Western blot analysis. ( B ) The mRNA level of COX-2 was detected by qRT-PCR. Saline: rats were injected with 100 μl saline only; Saline+SIN: rats were injected with 30 mg/kg SIN and 100 μl saline; CFA: rats were injected with 100 μl CFA only; CFA+SIN: rats were injected with 30 mg/kg SIN and 100 μl CFA. * p

Techniques Used: Expressing, Western Blot, Quantitative RT-PCR, Injection

37) Product Images from "Estrogen affects the negative feedback loop of PTENP1-miR200c to inhibit PTEN expression in the development of endometrioid endometrial carcinoma"

Article Title: Estrogen affects the negative feedback loop of PTENP1-miR200c to inhibit PTEN expression in the development of endometrioid endometrial carcinoma

Journal: Cell Death & Disease

doi: 10.1038/s41419-018-1207-4

Inverse expression of PTEN or PTENP1 and miR-200c in EEC tissue samples and cell lines. a In 37 of 40 EEC samples (92.5%), miR-200c was expressed at higher levels than that in adjacent normal tissues. b In 25 of 37 EEC samples (67.6%), PTEN was expressed at lower levels than in adjacent normal tissues. c In 25 of 27 EEC samples (92.6%), PTEN was expressed at lower levels than in adjacent normal tissues. d miR-200c, PTEN, and PTENP1 expression in 40 cases were analyzed, and showed there was a statistically correlation between their expression. e Immunohistochemistry showed that PTEN was lost in most complex atypical hyperplasia and cancer tissues. f GEO database was used to analyze the expression of miR-200c in EEC. g The expression of PTEN in 546 cases of endometrial carcinoma were analyzed in TGCA database by http://ualcan.path.uab.edu/analysis.html . h TGCA database was used to analyze the PTENP1 expression. i – j In four EEC cell lines the expression of PTEN, PTENP1, and miR-200c was detected by qRT-PCR and western blot. There was an opposite relationship between miR-200c and PTENP1/ PTEN. ** P
Figure Legend Snippet: Inverse expression of PTEN or PTENP1 and miR-200c in EEC tissue samples and cell lines. a In 37 of 40 EEC samples (92.5%), miR-200c was expressed at higher levels than that in adjacent normal tissues. b In 25 of 37 EEC samples (67.6%), PTEN was expressed at lower levels than in adjacent normal tissues. c In 25 of 27 EEC samples (92.6%), PTEN was expressed at lower levels than in adjacent normal tissues. d miR-200c, PTEN, and PTENP1 expression in 40 cases were analyzed, and showed there was a statistically correlation between their expression. e Immunohistochemistry showed that PTEN was lost in most complex atypical hyperplasia and cancer tissues. f GEO database was used to analyze the expression of miR-200c in EEC. g The expression of PTEN in 546 cases of endometrial carcinoma were analyzed in TGCA database by http://ualcan.path.uab.edu/analysis.html . h TGCA database was used to analyze the PTENP1 expression. i – j In four EEC cell lines the expression of PTEN, PTENP1, and miR-200c was detected by qRT-PCR and western blot. There was an opposite relationship between miR-200c and PTENP1/ PTEN. ** P

Techniques Used: Expressing, Immunohistochemistry, Quantitative RT-PCR, Western Blot

38) Product Images from "A Novel Transcriptional Activator, tubX, Is Required for the Stability of Bacillus sphaericus Mosquitocidal Plasmid pBsph"

Article Title: A Novel Transcriptional Activator, tubX, Is Required for the Stability of Bacillus sphaericus Mosquitocidal Plasmid pBsph

Journal: Journal of Bacteriology

doi: 10.1128/JB.01855-14

Effect of expressing additional tubRZ -Bs on the stability of pBsph in the Δ tubX mutant. (A) qRT-PCR analysis showing the mRNA levels of tubZ -Bs in strains R tubRZ (Δ tubX mutant with the tubRZ -Bs operon in the chromosome) and B tubRZ (Δ
Figure Legend Snippet: Effect of expressing additional tubRZ -Bs on the stability of pBsph in the Δ tubX mutant. (A) qRT-PCR analysis showing the mRNA levels of tubZ -Bs in strains R tubRZ (Δ tubX mutant with the tubRZ -Bs operon in the chromosome) and B tubRZ (Δ

Techniques Used: Expressing, Mutagenesis, Quantitative RT-PCR

qRT-PCR analysis of the mRNA abundances of tubX (A) and tubZ -Bs (B) relative to that of the rpoB reference in the Δ tubX mutant and its complemented strains. Total RNA was isolated from the cells of B. sphaericus strains grown to an OD 600 of 0.8.
Figure Legend Snippet: qRT-PCR analysis of the mRNA abundances of tubX (A) and tubZ -Bs (B) relative to that of the rpoB reference in the Δ tubX mutant and its complemented strains. Total RNA was isolated from the cells of B. sphaericus strains grown to an OD 600 of 0.8.

Techniques Used: Quantitative RT-PCR, Mutagenesis, Isolation

qRT-PCR analysis of the relative mRNA levels of tubZ -Bs in strains BH101 (G725 with tubRZ -Bs), BH102 (G725 with tubZ -Bs only), BH105 (G725 with tubRZX ), and BH106 (G725 with tubZX ). Means and standard deviations from at least four independent RNA samples
Figure Legend Snippet: qRT-PCR analysis of the relative mRNA levels of tubZ -Bs in strains BH101 (G725 with tubRZ -Bs), BH102 (G725 with tubZ -Bs only), BH105 (G725 with tubRZX ), and BH106 (G725 with tubZX ). Means and standard deviations from at least four independent RNA samples

Techniques Used: Quantitative RT-PCR

39) Product Images from "Galectins, Eosinophiles, and Macrophages May Contribute to Schistosoma japonicum Egg-Induced Immunopathology in a Mouse Model"

Article Title: Galectins, Eosinophiles, and Macrophages May Contribute to Schistosoma japonicum Egg-Induced Immunopathology in a Mouse Model

Journal: Frontiers in Immunology

doi: 10.3389/fimmu.2020.00146

Immunohistochemistry for CD68 + macrophages (A) , quantitative analysis of the positive areas (B) , and mRNA expressions of CD86, CD200R, IL-1β, iNOS, Arg1, and Ym1 (C) in the livers, spleens, and large intestines of S. japonicum -infected mice at 8 weeks p.i. Original magnification 400× (scale bar = 100 μm). There were four mice for immunohistochemistry analysis. For mRNA expression analysis, values are means from triplicate measurements and data are presented as means ± SD; there were eight mice in each group and the data represents from two experiments. * P
Figure Legend Snippet: Immunohistochemistry for CD68 + macrophages (A) , quantitative analysis of the positive areas (B) , and mRNA expressions of CD86, CD200R, IL-1β, iNOS, Arg1, and Ym1 (C) in the livers, spleens, and large intestines of S. japonicum -infected mice at 8 weeks p.i. Original magnification 400× (scale bar = 100 μm). There were four mice for immunohistochemistry analysis. For mRNA expression analysis, values are means from triplicate measurements and data are presented as means ± SD; there were eight mice in each group and the data represents from two experiments. * P

Techniques Used: Immunohistochemistry, Infection, Mouse Assay, Expressing

The ratios of M2/M1 gene expression (CD200R/CD86, Ym1/IL-1β, and Ym1/iNOS) in the livers, spleens, large intestines, and peritoneal macrophages of S. japonicum -infected mice at 8 weeks p.i. Data are presented as means ± SD; there were eight mice in each group and the data represents from two experiments. * P
Figure Legend Snippet: The ratios of M2/M1 gene expression (CD200R/CD86, Ym1/IL-1β, and Ym1/iNOS) in the livers, spleens, large intestines, and peritoneal macrophages of S. japonicum -infected mice at 8 weeks p.i. Data are presented as means ± SD; there were eight mice in each group and the data represents from two experiments. * P

Techniques Used: Expressing, Infection, Mouse Assay

The mRNA expressions of Gal-1, Gal-3, CD86, CD200R, IL-1β, iNOS, Arg1, and Ym1 in peritoneal macrophages of S. japonicum -infected mice at 8 weeks p.i. Values are means from triplicate measurements and data are presented as means ± SD; there were eight mice in each group and the data represents from two experiments. * P
Figure Legend Snippet: The mRNA expressions of Gal-1, Gal-3, CD86, CD200R, IL-1β, iNOS, Arg1, and Ym1 in peritoneal macrophages of S. japonicum -infected mice at 8 weeks p.i. Values are means from triplicate measurements and data are presented as means ± SD; there were eight mice in each group and the data represents from two experiments. * P

Techniques Used: Infection, Mouse Assay

40) Product Images from "Triggering unfolded protein response by 2-Deoxy-d-glucose inhibits porcine epidemic diarrhea virus propagation"

Article Title: Triggering unfolded protein response by 2-Deoxy-d-glucose inhibits porcine epidemic diarrhea virus propagation

Journal: Antiviral Research

doi: 10.1016/j.antiviral.2014.03.007

2-DG inhibits PEDV replication and gene expression. (A) Vero cells were treated with 5 mM 2-DG for the indicated time. Cell lysates were analyzed by Western blot using antibody against GRP78. (B) Vero cells were infected with PEDV (MOI = 0.01) after pretreated with 2-DG 24 h for indicated concentrations. Supernatant (virions) and cell lysates were analyzed by Western blot using antibody against PEDV N. (C) 2-DG affected PEDV replication. The cells were incubated either in the presence or absence of 2-DG for 24 h before infected with PEDV (MOI = 1). The cells were harvested for the indicated time for RT-PCR analysis using primers specific for PEDV N gene. The ratio was the gene copy number of N gene in 2-DG treated and mock-treated cells. (D) Vero cells were infected with PEDV (MOI = 0.01) for 48 h after cells treated with various dosages of 2-DG for 24 h. PEDV titers in the supernatants were measured by plaque formation assays. (E, F) 5 × 10 5 Vero cells were infected with PEDV (MOI = 0.01) for 48 h after cells treated with various dosages of 2-DG for 24 h. Total RNA was extracted, and cDNA was synthesized by reverse transcription. The RT-PCR was used to analyze the viral RNA copy numbers in the cells and supernatants (virion).
Figure Legend Snippet: 2-DG inhibits PEDV replication and gene expression. (A) Vero cells were treated with 5 mM 2-DG for the indicated time. Cell lysates were analyzed by Western blot using antibody against GRP78. (B) Vero cells were infected with PEDV (MOI = 0.01) after pretreated with 2-DG 24 h for indicated concentrations. Supernatant (virions) and cell lysates were analyzed by Western blot using antibody against PEDV N. (C) 2-DG affected PEDV replication. The cells were incubated either in the presence or absence of 2-DG for 24 h before infected with PEDV (MOI = 1). The cells were harvested for the indicated time for RT-PCR analysis using primers specific for PEDV N gene. The ratio was the gene copy number of N gene in 2-DG treated and mock-treated cells. (D) Vero cells were infected with PEDV (MOI = 0.01) for 48 h after cells treated with various dosages of 2-DG for 24 h. PEDV titers in the supernatants were measured by plaque formation assays. (E, F) 5 × 10 5 Vero cells were infected with PEDV (MOI = 0.01) for 48 h after cells treated with various dosages of 2-DG for 24 h. Total RNA was extracted, and cDNA was synthesized by reverse transcription. The RT-PCR was used to analyze the viral RNA copy numbers in the cells and supernatants (virion).

Techniques Used: Expressing, Western Blot, Infection, Incubation, Reverse Transcription Polymerase Chain Reaction, Synthesized

2-DG treatment affects virus packaging. (A) Vero cells were pretreated with 10 mM 2-DG for 24 h, and the cells were analyzed by FACS analysis using anti-CD13 polyclonal antibody. (B, C) 2-DG treatment did not affect virus entry as determined by RT-PCR and Western blot analysis. For RT-PCR analysis, the cells were treated with 10 mM 2-DG before virus infection. Bars represent mean percent copy number of viral RNA levels in cells by RT-PCR using primers specific for PEDV N gene. Cell lysates were analyzed by Western blot using antibody against PEDV N. (D) The Vero cells were infected with PEDV for 2 h (MOI = 1), 2-DG (10 mM) was incubated with the cells. The cells were harvested at different time points. Cell lysates were analyzed by Western blot using antibody against PEDV N. (E, F) The Vero cells was treated with different concentrations 2-DG after infected with PEDV (MOI = 0.01) for 2 h, the ratio of PEDV RNA copy numbers between the supernatants and the cell lysates was detected by Q-PCR, and the ratio of the virus titers in supernatants and cell lysates were determined by plaque assay.
Figure Legend Snippet: 2-DG treatment affects virus packaging. (A) Vero cells were pretreated with 10 mM 2-DG for 24 h, and the cells were analyzed by FACS analysis using anti-CD13 polyclonal antibody. (B, C) 2-DG treatment did not affect virus entry as determined by RT-PCR and Western blot analysis. For RT-PCR analysis, the cells were treated with 10 mM 2-DG before virus infection. Bars represent mean percent copy number of viral RNA levels in cells by RT-PCR using primers specific for PEDV N gene. Cell lysates were analyzed by Western blot using antibody against PEDV N. (D) The Vero cells were infected with PEDV for 2 h (MOI = 1), 2-DG (10 mM) was incubated with the cells. The cells were harvested at different time points. Cell lysates were analyzed by Western blot using antibody against PEDV N. (E, F) The Vero cells was treated with different concentrations 2-DG after infected with PEDV (MOI = 0.01) for 2 h, the ratio of PEDV RNA copy numbers between the supernatants and the cell lysates was detected by Q-PCR, and the ratio of the virus titers in supernatants and cell lysates were determined by plaque assay.

Techniques Used: FACS, Reverse Transcription Polymerase Chain Reaction, Western Blot, Infection, Incubation, Polymerase Chain Reaction, Plaque Assay

The impairment of UPR increased PEDV replication. (A) Vero cells were transfected with shRNAs to knockdown protein PERK for 48 h, and then infected with PEDV for 36 h. Scramble shRNA plasmid was used as the negative control. PERK, eIF2α and phos-eIF2α expression and the N protein of PEDV were analyzed by Western blot. β-Actin was used as sample loading control. (B) PEDV titers in the supernatants were measured by plaque formation assays. (C) The RNA level of gene ORF3 from the infected cells was evaluated by Q-PCR. (D, E) Vero cells were infected with PEDV for 2 h, then treated with 4-PBA for 24 h. The Western blot was performed to test eIF2α and phospho-eIF2α expression and the change of N protein of PEDV. β-Actin was used as sample loading control.
Figure Legend Snippet: The impairment of UPR increased PEDV replication. (A) Vero cells were transfected with shRNAs to knockdown protein PERK for 48 h, and then infected with PEDV for 36 h. Scramble shRNA plasmid was used as the negative control. PERK, eIF2α and phos-eIF2α expression and the N protein of PEDV were analyzed by Western blot. β-Actin was used as sample loading control. (B) PEDV titers in the supernatants were measured by plaque formation assays. (C) The RNA level of gene ORF3 from the infected cells was evaluated by Q-PCR. (D, E) Vero cells were infected with PEDV for 2 h, then treated with 4-PBA for 24 h. The Western blot was performed to test eIF2α and phospho-eIF2α expression and the change of N protein of PEDV. β-Actin was used as sample loading control.

Techniques Used: Transfection, Infection, shRNA, Plasmid Preparation, Negative Control, Expressing, Western Blot, Polymerase Chain Reaction

The transmission electron microscopy analysis of PEDV-infected cells. Vero cells were mock infected or infected with PEDV at MOI of 0.1 for 24 h. The cells were fixed and processed for electron microscopy analysis. (A) Mock infected Vero cells. (B) PEDV-infected Vero cells. The arrow indicated the swollen endoplasmic reticulum. (C) The swollen endoplasmic reticulum closed to nuclei in infected cells. (D) The vesicles full of virions.
Figure Legend Snippet: The transmission electron microscopy analysis of PEDV-infected cells. Vero cells were mock infected or infected with PEDV at MOI of 0.1 for 24 h. The cells were fixed and processed for electron microscopy analysis. (A) Mock infected Vero cells. (B) PEDV-infected Vero cells. The arrow indicated the swollen endoplasmic reticulum. (C) The swollen endoplasmic reticulum closed to nuclei in infected cells. (D) The vesicles full of virions.

Techniques Used: Transmission Assay, Electron Microscopy, Infection

PEDV induces the host cellular UPR signaling pathways. Vero cells were infected with PEDV at MOI of 0.01 or treated with Tu, the samples were collected at 0, 6, 12, 24, 36 and 48 h.p.i. (A–C) The expression of GRP78, PERK and eIF2α in the samples was analyzed by Western blot. β-Actin was used as sample loading control. (D) The cleavage of ATF6 during UPR was analyzed by Western blot. (E) Unspliced (uXBP-1) and spliced (sXBP-1) mRNA were analyzed by RT-PCR using specific primer pairs.
Figure Legend Snippet: PEDV induces the host cellular UPR signaling pathways. Vero cells were infected with PEDV at MOI of 0.01 or treated with Tu, the samples were collected at 0, 6, 12, 24, 36 and 48 h.p.i. (A–C) The expression of GRP78, PERK and eIF2α in the samples was analyzed by Western blot. β-Actin was used as sample loading control. (D) The cleavage of ATF6 during UPR was analyzed by Western blot. (E) Unspliced (uXBP-1) and spliced (sXBP-1) mRNA were analyzed by RT-PCR using specific primer pairs.

Techniques Used: Infection, Expressing, Western Blot, Reverse Transcription Polymerase Chain Reaction

PEDV replication is required for UPR induction. Vero cells were infected with UV-inactivated PEDV, PEDV (both MOI = 0.01) or treated with Tu (2 μg/ml). The cells were harvested at 24 h and Western blot was performed to determine UPR activation using anti-GRP78 antibody. β-Actin was used as a loading control.
Figure Legend Snippet: PEDV replication is required for UPR induction. Vero cells were infected with UV-inactivated PEDV, PEDV (both MOI = 0.01) or treated with Tu (2 μg/ml). The cells were harvested at 24 h and Western blot was performed to determine UPR activation using anti-GRP78 antibody. β-Actin was used as a loading control.

Techniques Used: Infection, Western Blot, Activation Assay

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RNA Extraction:

Article Title: Genome-Wide Identification and Analysis of Polygalacturonase Genes in Solanum lycopersicum
Article Snippet: .. Expression Analysis of SlPG Genes by qRT-PCR Total RNA was extracted using MiniBEST Plant RNA Extraction Kit (Takara, Japan). .. The first strand cDNA was prepared using Primer Script RT reagent kit (Takara, Japan).

Amplification:

Article Title: The potential contribution of miRNA-200-3p to the fatty acid metabolism by regulating AjEHHADH during aestivation in sea cucumber
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In Vitro:

Article Title: Shrimp Antiviral mja-miR-35 Targets CHI3L1 in Human M2 Macrophages and Suppresses Breast Cancer Metastasis
Article Snippet: .. The miRNAs and siRNAs were synthesized using in vitro transcription T7 kit (Takara) according to the manufacturer′s instructions. .. At different time after injection, the shrimp hemocytes were collected for later use.

Synthesized:

Article Title: CircPSMC3 suppresses the proliferation and metastasis of gastric cancer by acting as a competitive endogenous RNA through sponging miR-296-5p
Article Snippet: .. For circRNA and mRNA, cDNA was synthesized by using reverse transcription kit (Takara, Otsu, Japan) and for miRNA, total RNAs were reversed using RiboBio reverse transcription kit (Guangzhou, China). .. Quantification of mRNA and circular RNA was performed by using a SYBR Green PCR Kit (Takara, Otsu, Japan), and miRNA PCR was performed by using a SYBR Green PCR Kit (RiboBio,Guangzhou, China).

Article Title: Shrimp Antiviral mja-miR-35 Targets CHI3L1 in Human M2 Macrophages and Suppresses Breast Cancer Metastasis
Article Snippet: .. The miRNAs and siRNAs were synthesized using in vitro transcription T7 kit (Takara) according to the manufacturer′s instructions. .. At different time after injection, the shrimp hemocytes were collected for later use.

Quantitative RT-PCR:

Article Title: RNA-seq profiling reveals differentially expressed genes as potential markers for vital reaction in skin contusion: a pilot study
Article Snippet: .. Quantitative real-time reverse transcriptase polymerase chain reaction (RT-qPCR) Briefly, cDNA copies of total RNA were obtained using PrimerScript RT reagent Kit (TaKaRa, Japan). .. RT-qPCR reactions were run in 48-well reaction plates with an Illumina Eco Real-Time PCR System and a SYBR green kit (TaKaRa, Japan), according to the manufacturer's recommendations.

Article Title: Combined small RNA and gene expression analysis revealed roles of miRNAs in maize response to rice black-streaked dwarf virus infection
Article Snippet: .. qRT-PCR Validation For qRT-PCR validation of miRNAs, the Mir-X miRNA qRT-PCR SYBR Kit (Clontech Laboratories, Inc) were used following the manufacturer’s instructions. .. For all miRNAs and genes, the qRT-PCR was performed in ABI7500 (Applied Biosystems).

Article Title: Genome-Wide Identification and Analysis of Polygalacturonase Genes in Solanum lycopersicum
Article Snippet: .. Expression Analysis of SlPG Genes by qRT-PCR Total RNA was extracted using MiniBEST Plant RNA Extraction Kit (Takara, Japan). .. The first strand cDNA was prepared using Primer Script RT reagent kit (Takara, Japan).

Real-time Polymerase Chain Reaction:

Article Title: Circular RNA AKT3 upregulates PIK3R1 to enhance cisplatin resistance in gastric cancer via miR-198 suppression
Article Snippet: .. For Quantitative real-time PCR (RT-PCR), 500 ng of treated RNA was directly reverse transcribed using Prime Script RT Master Mix (Takara, Japan) and either random or oligo(dT) primers. .. Reverse transcription of miRNA was performed using a New Poly(A) Tailing Kit (ThermoFisher Scientific, China). mRNA was reverse transcribed into cDNA with a PrimeScript RT Master Mix Kit (Takara, RR036A, Japan). cDNA was amplified using Universal SYBR Green Master Mix (4,913,914,001, Roche, Shanghai, China).

Reverse Transcription Polymerase Chain Reaction:

Article Title: Circular RNA AKT3 upregulates PIK3R1 to enhance cisplatin resistance in gastric cancer via miR-198 suppression
Article Snippet: .. For Quantitative real-time PCR (RT-PCR), 500 ng of treated RNA was directly reverse transcribed using Prime Script RT Master Mix (Takara, Japan) and either random or oligo(dT) primers. .. Reverse transcription of miRNA was performed using a New Poly(A) Tailing Kit (ThermoFisher Scientific, China). mRNA was reverse transcribed into cDNA with a PrimeScript RT Master Mix Kit (Takara, RR036A, Japan). cDNA was amplified using Universal SYBR Green Master Mix (4,913,914,001, Roche, Shanghai, China).

Article Title: Development of a Conventional RT-PCR Assay for Rapid Detection of Porcine Deltacoronavirus with the Same Detection Limit as a SYBR Green-Based Real-Time RT-PCR Assay
Article Snippet: .. RT-PCR After isolating total RNA from samples, it was reverse transcribed into cDNA by following the manufacturer's instructions in PrimeScript™ 1st Strand cDNA Synthesis Kit (Takara Biotechnology, Dalian, China). .. Using Premix Taq™ kit, the PCR assay was performed under the following conditions: 1 μ L each primer (10 μ m), 25 μ L premix, 1 μ L cDNA, and 22 μ L distilled water.

Expressing:

Article Title: Genome-Wide Identification and Analysis of Polygalacturonase Genes in Solanum lycopersicum
Article Snippet: .. Expression Analysis of SlPG Genes by qRT-PCR Total RNA was extracted using MiniBEST Plant RNA Extraction Kit (Takara, Japan). .. The first strand cDNA was prepared using Primer Script RT reagent kit (Takara, Japan).

Article Title: The potential contribution of miRNA-200-3p to the fatty acid metabolism by regulating AjEHHADH during aestivation in sea cucumber
Article Snippet: .. Gene expression levels of AjEHHADH were analyzed using SYBR® Premix Ex Taq™ master mix (Cat No. RR420; Takara, Kusatsu, Japan) and amplification was measured by the StepOnePlus system (ABI Inc., Foster, CA, USA). .. Melting-curve analysis of the amplified products was performed at the end of each PCR reaction to ensure that only one PCR product was produced.

Polymerase Chain Reaction:

Article Title: RNA-seq profiling reveals differentially expressed genes as potential markers for vital reaction in skin contusion: a pilot study
Article Snippet: .. Quantitative real-time reverse transcriptase polymerase chain reaction (RT-qPCR) Briefly, cDNA copies of total RNA were obtained using PrimerScript RT reagent Kit (TaKaRa, Japan). .. RT-qPCR reactions were run in 48-well reaction plates with an Illumina Eco Real-Time PCR System and a SYBR green kit (TaKaRa, Japan), according to the manufacturer's recommendations.

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    TaKaRa sybr green qrt pcr master mix
    PTEN is a direct target gene of miR-130b. ( A ) PTEN was detected by <t>qRT-</t> <t>PCR</t> in 29 BC tissues and was matched the adjacent noncancerous breast tissues. ( B ) The relationship between PTEN and miR-130b expression was explored by Spearman’s correlation in 29 paired BC tissues and adjacent noncancerous breast tissues. PTEN was inversely correlated with the miR-130b levels in BC tissues. ( C ) Relative PTEN expression in MCF-7 and MCF-7/ADR cells were detected by qRT-PCR (left panel) and western blot (right panel). ( D ) Effects of miR-130b mimics in MCF-7 cells and miR-130b inhibitor in MCF-7/ADR cells on the relative expression levels of PTEN were detected by qRT- PCR (left panel) and western blot (right panel). ( E ) PTEN expression was detected by immunofluorescence staining in these cells. Red fluorescence: PTEN, blue fluorescence: DAPI, staining for nuclear DNA. There were no significant differences between the control group and the blank group. ( F ) The relative luciferase activity of 293 T cells was detected after the pGL3-control vector, wt or mut PTEN 3′UTR genes were co-transfected with miR-NC or miR-130b mimics (*P
    Sybr Green Qrt Pcr Master Mix, supplied by TaKaRa, used in various techniques. Bioz Stars score: 89/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Expression of screened <t>miRNAs</t> in serum exosomes of STZ treated mice. Serum was sampled from ICR mice at different time after STZ injection. Fasting tail blood glucose ( A ) and serum insulin ( B ) levels were measured. The expression of 4 serum exosomal miRNAs selected from microarray miR-375-3p ( C ), miR-129-5p ( D ), miR-378a-3p ( E ), miR-382-5p ( F ) were analyzed using <t>qRT-PCR.</t> Values represent means ± SEM (n≥9 mice of each time point). (* P
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    TaKaRa sybr real time qrt pcr
    TIMP-3 3’-UTR is a direct target for miR-103. (A) Suppression of miR-103 by anti-miR-103 in Ishikawa cells is shown. The cells were frst transfected with anti-miR-103 or negative control oligonucleotide. The cells were harvested 2 days later and total RNA was extracted using TRIzol reagent. Mature miR-103 was detected by <t>SYBR</t> <t>qRT-PCR,</t> using U6 RNA for normalization. (B) Suppression of miR-103 by anti-miR-103 in HEC-1B cells is shown. Following transfection with anti-miR-103 (Anti-miR) or negative control oligonucleotide (Neg Control), mature miR-103 was detected as in (A). (C) An evolutionarily conserved target sequence for miR-103 is detected in the 3’-UTR of TIMP-3. (D and E) The luciferase assay was performed using Ishikawa and HEC-1B cells, respectively. The effect of miR-103 on the luciferase activity of pGL3-TIMP-3-wt and pGL3-TIMP-3-mut was measured as described in Materials and methods. A statistically significant upregulation in luciferase activity was found when the cells were transfected with pGL3-TIMP-3-wt together with anti-miR-103. pGL3-TIMP-3-wt, vector with wild-type TIMP-3 3’-UTR; pGL3-TIMP-3-mut, vector with mutated TIMP-3 3’-UTR. Each bar is the mean ± SEM from three independent experiments. * P
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    PTEN is a direct target gene of miR-130b. ( A ) PTEN was detected by qRT- PCR in 29 BC tissues and was matched the adjacent noncancerous breast tissues. ( B ) The relationship between PTEN and miR-130b expression was explored by Spearman’s correlation in 29 paired BC tissues and adjacent noncancerous breast tissues. PTEN was inversely correlated with the miR-130b levels in BC tissues. ( C ) Relative PTEN expression in MCF-7 and MCF-7/ADR cells were detected by qRT-PCR (left panel) and western blot (right panel). ( D ) Effects of miR-130b mimics in MCF-7 cells and miR-130b inhibitor in MCF-7/ADR cells on the relative expression levels of PTEN were detected by qRT- PCR (left panel) and western blot (right panel). ( E ) PTEN expression was detected by immunofluorescence staining in these cells. Red fluorescence: PTEN, blue fluorescence: DAPI, staining for nuclear DNA. There were no significant differences between the control group and the blank group. ( F ) The relative luciferase activity of 293 T cells was detected after the pGL3-control vector, wt or mut PTEN 3′UTR genes were co-transfected with miR-NC or miR-130b mimics (*P

    Journal: Scientific Reports

    Article Title: MicroRNA-130b targets PTEN to mediate drug resistance and proliferation of breast cancer cells via the PI3K/Akt signaling pathway

    doi: 10.1038/srep41942

    Figure Lengend Snippet: PTEN is a direct target gene of miR-130b. ( A ) PTEN was detected by qRT- PCR in 29 BC tissues and was matched the adjacent noncancerous breast tissues. ( B ) The relationship between PTEN and miR-130b expression was explored by Spearman’s correlation in 29 paired BC tissues and adjacent noncancerous breast tissues. PTEN was inversely correlated with the miR-130b levels in BC tissues. ( C ) Relative PTEN expression in MCF-7 and MCF-7/ADR cells were detected by qRT-PCR (left panel) and western blot (right panel). ( D ) Effects of miR-130b mimics in MCF-7 cells and miR-130b inhibitor in MCF-7/ADR cells on the relative expression levels of PTEN were detected by qRT- PCR (left panel) and western blot (right panel). ( E ) PTEN expression was detected by immunofluorescence staining in these cells. Red fluorescence: PTEN, blue fluorescence: DAPI, staining for nuclear DNA. There were no significant differences between the control group and the blank group. ( F ) The relative luciferase activity of 293 T cells was detected after the pGL3-control vector, wt or mut PTEN 3′UTR genes were co-transfected with miR-NC or miR-130b mimics (*P

    Article Snippet: The qRT-PCR for the analysis of PTEN mRNA expression was performed using the SYBR Green qRT-PCR master mix (TaKaRa, Otsu, Shiga, Japan) and GAPDH (glyceraldehyde-3-phosphate dehydrogenase) as an internal control.

    Techniques: Quantitative RT-PCR, Expressing, Western Blot, Immunofluorescence, Staining, Fluorescence, Luciferase, Activity Assay, Plasmid Preparation, Transfection

    Chondrocyte marker gene expression analyses of the single subclone derived from mixed piCLCs (referred to as piCLCs-S). a Cell morphologies of piCLCs-S. b Toluidine blue staining and alcian blue staining of piCLCs-S. c, d qRT-PCR analysis of chondrocyte ( c ) and fibroblast ( d ) marker gene expression in the indicated cells. In ( c ) and ( d ), error bars indicate mean ± SD ( n = 3). e Immunofluorescence analysis showing protein expression of type I collagen (type I C), type II collagen (type II C) and aggrecan in the indicated cells. Nuclei were counterstained with DAPI

    Journal: Cell Death Discovery

    Article Title: Direct conversion of pig fibroblasts to chondrocyte-like cells by c-Myc

    doi: 10.1038/s41420-018-0136-4

    Figure Lengend Snippet: Chondrocyte marker gene expression analyses of the single subclone derived from mixed piCLCs (referred to as piCLCs-S). a Cell morphologies of piCLCs-S. b Toluidine blue staining and alcian blue staining of piCLCs-S. c, d qRT-PCR analysis of chondrocyte ( c ) and fibroblast ( d ) marker gene expression in the indicated cells. In ( c ) and ( d ), error bars indicate mean ± SD ( n = 3). e Immunofluorescence analysis showing protein expression of type I collagen (type I C), type II collagen (type II C) and aggrecan in the indicated cells. Nuclei were counterstained with DAPI

    Article Snippet: Total RNA was reversely transcribed with the PrimeScript RT reagent Kit (TaKaRa). qRT-PCR for the expression of mRNA analysis was performed using SYBR Green qRT-PCR master mix (TaKaRa) on a Stratagene Mx3005P qRT-PCR System (Agilent Technologies, Santa Clara, CA).

    Techniques: Marker, Expressing, Derivative Assay, Staining, Quantitative RT-PCR, Immunofluorescence

    Chondrocyte marker gene expression analyses of pig induced chondrocyte-like cells (piCLCs) from PEFs by c-Myc. a Cell morphologies of PEFs, pPr Ch and piCLCs. The phase-contrast photographs of c-Myc-expressing PEFs (i.e., piCLCs) and vector-expressing PEFs were token at day 4 post-infection. pPr Ch: porcine primary chondrocytes. b Toluidine blue staining and alcian blue staining for piCLCs. Proteoglycan was identified by toluidine blue staining and alcian blue staining. c , d qRT-PCR analysis of chondrocyte ( c ) and fibroblast ( d ) marker gene expression in the indicated cells. In ( c ) and ( d ), error bars indicate mean ± SD ( n = 3). e Cell extracts from pPr Ch, piCLCs and PEFs were analysed by immunoblotting with antibodies against the indicated proteins. Lane 1: pPr Ch; Lane 2: piCLCs P7; Lane 3: piCLCs P14; Lane 4: piCLCs P19; Lane 5: piCLCs P25; Lane 6: piCLCs-S; Lane 7: PEFs. f Immunofluorescence analysis showing protein expression of type I collagen (type I C), type II collagen (type II C) and aggrecan in the indicated cells. Nuclei were counterstained with DAPI

    Journal: Cell Death Discovery

    Article Title: Direct conversion of pig fibroblasts to chondrocyte-like cells by c-Myc

    doi: 10.1038/s41420-018-0136-4

    Figure Lengend Snippet: Chondrocyte marker gene expression analyses of pig induced chondrocyte-like cells (piCLCs) from PEFs by c-Myc. a Cell morphologies of PEFs, pPr Ch and piCLCs. The phase-contrast photographs of c-Myc-expressing PEFs (i.e., piCLCs) and vector-expressing PEFs were token at day 4 post-infection. pPr Ch: porcine primary chondrocytes. b Toluidine blue staining and alcian blue staining for piCLCs. Proteoglycan was identified by toluidine blue staining and alcian blue staining. c , d qRT-PCR analysis of chondrocyte ( c ) and fibroblast ( d ) marker gene expression in the indicated cells. In ( c ) and ( d ), error bars indicate mean ± SD ( n = 3). e Cell extracts from pPr Ch, piCLCs and PEFs were analysed by immunoblotting with antibodies against the indicated proteins. Lane 1: pPr Ch; Lane 2: piCLCs P7; Lane 3: piCLCs P14; Lane 4: piCLCs P19; Lane 5: piCLCs P25; Lane 6: piCLCs-S; Lane 7: PEFs. f Immunofluorescence analysis showing protein expression of type I collagen (type I C), type II collagen (type II C) and aggrecan in the indicated cells. Nuclei were counterstained with DAPI

    Article Snippet: Total RNA was reversely transcribed with the PrimeScript RT reagent Kit (TaKaRa). qRT-PCR for the expression of mRNA analysis was performed using SYBR Green qRT-PCR master mix (TaKaRa) on a Stratagene Mx3005P qRT-PCR System (Agilent Technologies, Santa Clara, CA).

    Techniques: Marker, Expressing, Plasmid Preparation, Infection, Staining, Quantitative RT-PCR, Immunofluorescence

    Expression of screened miRNAs in serum exosomes of STZ treated mice. Serum was sampled from ICR mice at different time after STZ injection. Fasting tail blood glucose ( A ) and serum insulin ( B ) levels were measured. The expression of 4 serum exosomal miRNAs selected from microarray miR-375-3p ( C ), miR-129-5p ( D ), miR-378a-3p ( E ), miR-382-5p ( F ) were analyzed using qRT-PCR. Values represent means ± SEM (n≥9 mice of each time point). (* P

    Journal: Aging (Albany NY)

    Article Title: Injury factors alter miRNAs profiles of exosomes derived from islets and circulation

    doi: 10.18632/aging.101689

    Figure Lengend Snippet: Expression of screened miRNAs in serum exosomes of STZ treated mice. Serum was sampled from ICR mice at different time after STZ injection. Fasting tail blood glucose ( A ) and serum insulin ( B ) levels were measured. The expression of 4 serum exosomal miRNAs selected from microarray miR-375-3p ( C ), miR-129-5p ( D ), miR-378a-3p ( E ), miR-382-5p ( F ) were analyzed using qRT-PCR. Values represent means ± SEM (n≥9 mice of each time point). (* P

    Article Snippet: Expression of miRNA was analyzed using MiR-X miRNA qRT-PCR SYBR Kit and One Step SYBR PrimeScript RT-PCR Kit, respectively (Clontech Laboratories, Inc., Mountain View, USA).

    Techniques: Expressing, Mouse Assay, Injection, Microarray, Quantitative RT-PCR

    Validation of exosomal miRNAs detected by microarray. Four miRNAs with top high FC values (FC > 1.5 in both STZ and cytokines treated groups) miR-375-3p ( A ), miR-129-5p ( B ), miR-378a-3p ( C ), miR-382-5p ( D ) were further verified by qRT-PCR in three independent samples. The data are presented as mean±SEM. (* P

    Journal: Aging (Albany NY)

    Article Title: Injury factors alter miRNAs profiles of exosomes derived from islets and circulation

    doi: 10.18632/aging.101689

    Figure Lengend Snippet: Validation of exosomal miRNAs detected by microarray. Four miRNAs with top high FC values (FC > 1.5 in both STZ and cytokines treated groups) miR-375-3p ( A ), miR-129-5p ( B ), miR-378a-3p ( C ), miR-382-5p ( D ) were further verified by qRT-PCR in three independent samples. The data are presented as mean±SEM. (* P

    Article Snippet: Expression of miRNA was analyzed using MiR-X miRNA qRT-PCR SYBR Kit and One Step SYBR PrimeScript RT-PCR Kit, respectively (Clontech Laboratories, Inc., Mountain View, USA).

    Techniques: Microarray, Quantitative RT-PCR

    Serum exosomal miRNA-375 and miRNA-129 expression in T1DM and T2DM patients with different β cell function. After standard mixed meal tolerance tests, the area under curve (AUC) of plasma glucose were measured ( A ), and the ratio of C-peptide AUC to glucose AUC were calculated to evaluate islet β cell function ( B ). Serum exosomal miRNA-375 ( E , F ) and miRNA-129 ( G , H ) expression in T1DM and T2DM patients with diverse diabetes course were detected by qRT-PCR. All measurements are mean ± SEM (n≥10 subjects of each group) (* P

    Journal: Aging (Albany NY)

    Article Title: Injury factors alter miRNAs profiles of exosomes derived from islets and circulation

    doi: 10.18632/aging.101689

    Figure Lengend Snippet: Serum exosomal miRNA-375 and miRNA-129 expression in T1DM and T2DM patients with different β cell function. After standard mixed meal tolerance tests, the area under curve (AUC) of plasma glucose were measured ( A ), and the ratio of C-peptide AUC to glucose AUC were calculated to evaluate islet β cell function ( B ). Serum exosomal miRNA-375 ( E , F ) and miRNA-129 ( G , H ) expression in T1DM and T2DM patients with diverse diabetes course were detected by qRT-PCR. All measurements are mean ± SEM (n≥10 subjects of each group) (* P

    Article Snippet: Expression of miRNA was analyzed using MiR-X miRNA qRT-PCR SYBR Kit and One Step SYBR PrimeScript RT-PCR Kit, respectively (Clontech Laboratories, Inc., Mountain View, USA).

    Techniques: Expressing, Cell Function Assay, Quantitative RT-PCR

    TIMP-3 3’-UTR is a direct target for miR-103. (A) Suppression of miR-103 by anti-miR-103 in Ishikawa cells is shown. The cells were frst transfected with anti-miR-103 or negative control oligonucleotide. The cells were harvested 2 days later and total RNA was extracted using TRIzol reagent. Mature miR-103 was detected by SYBR qRT-PCR, using U6 RNA for normalization. (B) Suppression of miR-103 by anti-miR-103 in HEC-1B cells is shown. Following transfection with anti-miR-103 (Anti-miR) or negative control oligonucleotide (Neg Control), mature miR-103 was detected as in (A). (C) An evolutionarily conserved target sequence for miR-103 is detected in the 3’-UTR of TIMP-3. (D and E) The luciferase assay was performed using Ishikawa and HEC-1B cells, respectively. The effect of miR-103 on the luciferase activity of pGL3-TIMP-3-wt and pGL3-TIMP-3-mut was measured as described in Materials and methods. A statistically significant upregulation in luciferase activity was found when the cells were transfected with pGL3-TIMP-3-wt together with anti-miR-103. pGL3-TIMP-3-wt, vector with wild-type TIMP-3 3’-UTR; pGL3-TIMP-3-mut, vector with mutated TIMP-3 3’-UTR. Each bar is the mean ± SEM from three independent experiments. * P

    Journal: Oncology Letters

    Article Title: microRNA-103 regulates the growth and invasion of endometrial cancer cells through the downregulation of tissue inhibitor of metalloproteinase 3

    doi: 10.3892/ol.2012.638

    Figure Lengend Snippet: TIMP-3 3’-UTR is a direct target for miR-103. (A) Suppression of miR-103 by anti-miR-103 in Ishikawa cells is shown. The cells were frst transfected with anti-miR-103 or negative control oligonucleotide. The cells were harvested 2 days later and total RNA was extracted using TRIzol reagent. Mature miR-103 was detected by SYBR qRT-PCR, using U6 RNA for normalization. (B) Suppression of miR-103 by anti-miR-103 in HEC-1B cells is shown. Following transfection with anti-miR-103 (Anti-miR) or negative control oligonucleotide (Neg Control), mature miR-103 was detected as in (A). (C) An evolutionarily conserved target sequence for miR-103 is detected in the 3’-UTR of TIMP-3. (D and E) The luciferase assay was performed using Ishikawa and HEC-1B cells, respectively. The effect of miR-103 on the luciferase activity of pGL3-TIMP-3-wt and pGL3-TIMP-3-mut was measured as described in Materials and methods. A statistically significant upregulation in luciferase activity was found when the cells were transfected with pGL3-TIMP-3-wt together with anti-miR-103. pGL3-TIMP-3-wt, vector with wild-type TIMP-3 3’-UTR; pGL3-TIMP-3-mut, vector with mutated TIMP-3 3’-UTR. Each bar is the mean ± SEM from three independent experiments. * P

    Article Snippet: Briefly, miRNAs and mRNA were examined by SYBR real-time qRT-PCR (Takara, Dalian, China) on a 7500 Real-Time PCR system (Applied Biosystems, Carlsbad, CA, USA) according to the manufacturer’s instructions.

    Techniques: Transfection, Negative Control, Quantitative RT-PCR, Sequencing, Luciferase, Activity Assay, Plasmid Preparation