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Flowchart of <t>microarray</t> data analysis. Swine monocytes were exposed to Bb12 with or without the addition of a blocking antibody anti-TLR2. Groups were as follows: unstimulated swine monocytes, monocytes stimulated with Bb12 for 4 h, and swine monocytes incubated with anti-TLR2 antibody and Bb12 for 4 h. Microarray was performed, and 40 microRNAs with a MFI > 500 were selected for analysis of molecular interactions (KEGG analysis). The microRNAs were also analyzed for miRNA–mRNA interactions within TLR2 pathway targets (miRTarBase). After this, 15 miRNAs with a reported interaction with TLR2 pathway–related target proteins were selected to analyze with a multiple t test
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Flowchart of <t>microarray</t> data analysis. Swine monocytes were exposed to Bb12 with or without the addition of a blocking antibody anti-TLR2. Groups were as follows: unstimulated swine monocytes, monocytes stimulated with Bb12 for 4 h, and swine monocytes incubated with anti-TLR2 antibody and Bb12 for 4 h. Microarray was performed, and 40 microRNAs with a MFI > 500 were selected for analysis of molecular interactions (KEGG analysis). The microRNAs were also analyzed for miRNA–mRNA interactions within TLR2 pathway targets (miRTarBase). After this, 15 miRNAs with a reported interaction with TLR2 pathway–related target proteins were selected to analyze with a multiple t test
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Mir-92b promoted osteogenesis in MSCs (A–B) The top three up/down-regulated microRNAs in De-Os-MSCs were listed, and verified by qPCR (B) (C–E) The scrambled control, let-7e, mir-10b, mir-20a, mir-92b, mir-371 and mir-373 were transduced into MSCs with lentiviruses. The overexpression of each <t>microRNA</t> was verified by qPCR (C). The infected MSCs were induced to undergo osteogenic differentiation for 10 days, then the calcium deposits were stained with Alizarin Red S (D), and quantified (E) (F) Total RNA was extracted from MSCs infected with mir-92b or scrambled control. The mRNA expression levels of Osterix, Runx2, OPN and ALP were detected by qPCR. β-actin was used as an internal control. The data was expressed as mean ​± ​SD (n ​= ​3). ∗p ​< ​0.05 (G–I) Total proteins were extracted from MSCs transduced with scrambled control or mir-92b. Then the proteins were analyzed by western blot using indicated antibodies. The protein levels of pERK (H) and pJNK (I) was normalized to ERK and JNK1 respectively. All the data represent mean ​± ​SD of three independent experiments. ∗p ​< ​0.05 (J–K) The mir-92b antagmir was transfected into MSCs, then the cells were treated with osteogenic induction medium for 10 days, the calcium deposits were stained with Alizarin Red S (J), the changes of osteogenesis-related genes was checked by qPCR (K).
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Thermo Fisher mirnas microarray analysis microrna 4.0 array
Validation of candidate <t>miRNAs.</t> Among the top rated nine miRNAs screened from our miRNA <t>microarray,</t> four miRNAs involving (A) hsa-miR-145-5p, (B) hsa-miR-497-5p, (C) hsa-miR-29a-3p and (D) hsa-miR-204-5p were also significantly altered in GSE40355. miRNA, microRNA.
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Flowchart of microarray data analysis. Swine monocytes were exposed to Bb12 with or without the addition of a blocking antibody anti-TLR2. Groups were as follows: unstimulated swine monocytes, monocytes stimulated with Bb12 for 4 h, and swine monocytes incubated with anti-TLR2 antibody and Bb12 for 4 h. Microarray was performed, and 40 microRNAs with a MFI > 500 were selected for analysis of molecular interactions (KEGG analysis). The microRNAs were also analyzed for miRNA–mRNA interactions within TLR2 pathway targets (miRTarBase). After this, 15 miRNAs with a reported interaction with TLR2 pathway–related target proteins were selected to analyze with a multiple t test

Journal: Probiotics and Antimicrobial Proteins

Article Title: Immunomodulation by Bifidobacterium animalis subsp. lactis Bb12: Integrative Analysis of miRNA Expression and TLR2 Pathway–Related Target Proteins in Swine Monocytes

doi: 10.1007/s12602-021-09816-1

Figure Lengend Snippet: Flowchart of microarray data analysis. Swine monocytes were exposed to Bb12 with or without the addition of a blocking antibody anti-TLR2. Groups were as follows: unstimulated swine monocytes, monocytes stimulated with Bb12 for 4 h, and swine monocytes incubated with anti-TLR2 antibody and Bb12 for 4 h. Microarray was performed, and 40 microRNAs with a MFI > 500 were selected for analysis of molecular interactions (KEGG analysis). The microRNAs were also analyzed for miRNA–mRNA interactions within TLR2 pathway targets (miRTarBase). After this, 15 miRNAs with a reported interaction with TLR2 pathway–related target proteins were selected to analyze with a multiple t test

Article Snippet: A total of 2 μg RNA from each sample was sent for genome-wide microRNA microarray analysis using μParaflo® microfluidic biochip technology; this service was provided by LC Sciences (Houston, TX, USA).

Techniques: Microarray, Blocking Assay, Incubation

Comparison of qRT-PCR and microarray results. Log2 fold change expression between unstimulated monocytes and Bb12-stimulated cells by qRT-PCR and microarray

Journal: Probiotics and Antimicrobial Proteins

Article Title: Immunomodulation by Bifidobacterium animalis subsp. lactis Bb12: Integrative Analysis of miRNA Expression and TLR2 Pathway–Related Target Proteins in Swine Monocytes

doi: 10.1007/s12602-021-09816-1

Figure Lengend Snippet: Comparison of qRT-PCR and microarray results. Log2 fold change expression between unstimulated monocytes and Bb12-stimulated cells by qRT-PCR and microarray

Article Snippet: A total of 2 μg RNA from each sample was sent for genome-wide microRNA microarray analysis using μParaflo® microfluidic biochip technology; this service was provided by LC Sciences (Houston, TX, USA).

Techniques: Comparison, Quantitative RT-PCR, Microarray, Expressing

Mir-92b promoted osteogenesis in MSCs (A–B) The top three up/down-regulated microRNAs in De-Os-MSCs were listed, and verified by qPCR (B) (C–E) The scrambled control, let-7e, mir-10b, mir-20a, mir-92b, mir-371 and mir-373 were transduced into MSCs with lentiviruses. The overexpression of each microRNA was verified by qPCR (C). The infected MSCs were induced to undergo osteogenic differentiation for 10 days, then the calcium deposits were stained with Alizarin Red S (D), and quantified (E) (F) Total RNA was extracted from MSCs infected with mir-92b or scrambled control. The mRNA expression levels of Osterix, Runx2, OPN and ALP were detected by qPCR. β-actin was used as an internal control. The data was expressed as mean ​± ​SD (n ​= ​3). ∗p ​< ​0.05 (G–I) Total proteins were extracted from MSCs transduced with scrambled control or mir-92b. Then the proteins were analyzed by western blot using indicated antibodies. The protein levels of pERK (H) and pJNK (I) was normalized to ERK and JNK1 respectively. All the data represent mean ​± ​SD of three independent experiments. ∗p ​< ​0.05 (J–K) The mir-92b antagmir was transfected into MSCs, then the cells were treated with osteogenic induction medium for 10 days, the calcium deposits were stained with Alizarin Red S (J), the changes of osteogenesis-related genes was checked by qPCR (K).

Journal: Journal of Orthopaedic Translation

Article Title: De-osteogenic-differentiated mesenchymal stem cells accelerate fracture healing by mir-92b

doi: 10.1016/j.jot.2020.10.009

Figure Lengend Snippet: Mir-92b promoted osteogenesis in MSCs (A–B) The top three up/down-regulated microRNAs in De-Os-MSCs were listed, and verified by qPCR (B) (C–E) The scrambled control, let-7e, mir-10b, mir-20a, mir-92b, mir-371 and mir-373 were transduced into MSCs with lentiviruses. The overexpression of each microRNA was verified by qPCR (C). The infected MSCs were induced to undergo osteogenic differentiation for 10 days, then the calcium deposits were stained with Alizarin Red S (D), and quantified (E) (F) Total RNA was extracted from MSCs infected with mir-92b or scrambled control. The mRNA expression levels of Osterix, Runx2, OPN and ALP were detected by qPCR. β-actin was used as an internal control. The data was expressed as mean ​± ​SD (n ​= ​3). ∗p ​< ​0.05 (G–I) Total proteins were extracted from MSCs transduced with scrambled control or mir-92b. Then the proteins were analyzed by western blot using indicated antibodies. The protein levels of pERK (H) and pJNK (I) was normalized to ERK and JNK1 respectively. All the data represent mean ​± ​SD of three independent experiments. ∗p ​< ​0.05 (J–K) The mir-92b antagmir was transfected into MSCs, then the cells were treated with osteogenic induction medium for 10 days, the calcium deposits were stained with Alizarin Red S (J), the changes of osteogenesis-related genes was checked by qPCR (K).

Article Snippet: The microRNA microarray analysis was performed by the Annoroad Gene Technology Corporation (Beijing, China).

Techniques: Control, Over Expression, Infection, Staining, Expressing, Transduction, Western Blot, Transfection

Validation of candidate miRNAs. Among the top rated nine miRNAs screened from our miRNA microarray, four miRNAs involving (A) hsa-miR-145-5p, (B) hsa-miR-497-5p, (C) hsa-miR-29a-3p and (D) hsa-miR-204-5p were also significantly altered in GSE40355. miRNA, microRNA.

Journal: Molecular Medicine Reports

Article Title: Identification and bioinformatics analysis of miRNAs associated with human muscle invasive bladder cancer

doi: 10.3892/mmr.2017.7726

Figure Lengend Snippet: Validation of candidate miRNAs. Among the top rated nine miRNAs screened from our miRNA microarray, four miRNAs involving (A) hsa-miR-145-5p, (B) hsa-miR-497-5p, (C) hsa-miR-29a-3p and (D) hsa-miR-204-5p were also significantly altered in GSE40355. miRNA, microRNA.

Article Snippet: After assessing RNA quality and quantity, the miRNAs microarray analysis (Affymetrix microRNA 4.0 Array, Affymetrix, Inc., Santa Clara, CA, USA) was performed according to the manufacturer's instructions.

Techniques: Microarray