Journal: International Journal of Molecular Sciences

Article Title: Phospholipase A1 Member A Activates Fibroblast-like Synoviocytes through the Autotaxin-Lysophosphatidic Acid Receptor Axis

doi: 10.3390/ijms222312685

Figure Lengend Snippet: Presence of PLA1A in cultured human primary FLSs. ( A ) Total RNA from human primary FLSs from normal donors were tested for the expression of PLA1A by semi-quantitative RT-PCR. Two pairs PLA1A oligo primers were tested. GAPDH was used as a house-keeping gene. Total RNA from human testis tissue was used as a positive control. ( B ) The presence of PLA1A in human primary FLSs cultured in the absence or presence of 10% FBS were examined using western blot. GAPDH was used as an internal loading control. Recombinant PLA1A protein was used as a positive control. The blots shown are representative of five independent experiments with similar results. Amount of PLA1A was quantified densitometrically and was normalized with respect to total GAPDH. The data were the means ± SE from five experiments. ( C ) The presence of PLA1A in human primary FLSs cultured in the absence or presence of 10% FBS were examined using flow cytometry. Cells were permeabilized with digitonin, and rabbit IgG isotype was used as a control. The figures shown were representative of three independent experiments with similar results. The data were the means ± SE from three experiments. MFI, mean fluorescence intensity.

Article Snippet: Normal human primary FLSs were purchased from Asterand (Detroit, MI, USA) or provided by Dr. Di Battista (MUHC-Research Institute).

Techniques: Cell Culture, Expressing, Quantitative RT-PCR, Positive Control, Western Blot, Control, Recombinant, Flow Cytometry, Fluorescence

Journal: International Journal of Molecular Sciences

Article Title: Phospholipase A1 Member A Activates Fibroblast-like Synoviocytes through the Autotaxin-Lysophosphatidic Acid Receptor Axis

doi: 10.3390/ijms222312685

Figure Lengend Snippet: IL-8 production in human primary FLSs stimulated with PLA1A. Human primary FLSs from normal donors were stimulated with 0.2 µg/mL or 0.5 µg/mL recombinant PLA1A for 24 h. The amounts of IL-8 released in the supernatants were monitored using ELISA. IL-8 produced by FLSs cultured in serum-free DMEM was 4.03 ± 1.53 pg/mL, and for each experiment, basal IL-8 production was set at 100% for data normalization. The data shown are the means ± SE from three experiments. For statistical comparative analyses, cytokine amount in cells stimulated with PLA1A were compared to that in non-stimulated cells. ** p < 0.01.

Article Snippet: Normal human primary FLSs were purchased from Asterand (Detroit, MI, USA) or provided by Dr. Di Battista (MUHC-Research Institute).

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Produced, Cell Culture

Journal: International Journal of Molecular Sciences

Article Title: Phospholipase A1 Member A Activates Fibroblast-like Synoviocytes through the Autotaxin-Lysophosphatidic Acid Receptor Axis

doi: 10.3390/ijms222312685

Figure Lengend Snippet: Exposure of phosphatidylserine (PS) and effects of PLA1A on activated human primary FLSs. ( A ) Human primary FLSs were activated with 100 ng/mL TNF for 1, 5, 15, 30, and 60 min. Surface exposure of PS on collected cells were stained with propidium iodide (PI) and FITC Annexin V and examined using flow cytometry. FITC Annexin V-positive and PI-negative cells were considered as PS-positive viable cells. ( B ) Human primary FLSs were treated with 0.2 µg/mL recombinant PLA1A in the absence or presence of heparin at concentrations of 25, 50, 100, 200, 400, 800, and 1600 µg/mL for 24 h. Heparin was added to cell culture 30 min prior to PLA1A. The amounts of IL-8 released in the supernatants were monitored using ELISA. IL-8 produced by FLSs cultured in serum-free DMEM was 5.37 ± 1.43 pg/mL, and for each experiment, basal IL-8 production was set at 100% for data normalization. The data were the means ± SE from three experiments.

Article Snippet: Normal human primary FLSs were purchased from Asterand (Detroit, MI, USA) or provided by Dr. Di Battista (MUHC-Research Institute).

Techniques: Staining, Flow Cytometry, Recombinant, Cell Culture, Enzyme-linked Immunosorbent Assay, Produced

Journal: International Journal of Molecular Sciences

Article Title: Phospholipase A1 Member A Activates Fibroblast-like Synoviocytes through the Autotaxin-Lysophosphatidic Acid Receptor Axis

doi: 10.3390/ijms222312685

Figure Lengend Snippet: Pro-inflammatory effects of LPA, lysoPS, and PLA1A in human primary FLSs. ( A ) Total RNA from human primary FLSs from normal donors were tested for the expression of lysoPS receptors using semi-quantitative RT-PCR. For each lysoPS receptor (GRP34, GPR174, and P2RY10), three pairs oligo primers were tested. Data shown are results of one pair primer for each receptor. GAPDH was used as a house-keeping gene. Total RNA from human CD4 + T memory cells was used as a positive control. ( B – D ) Human primary FLSs from normal donors were treated with 5 µM LPA ( B ), or 5 µM lysoPS ( C ), or 0.2 µg/mL PLA1A ( D ) in the presence of 5 µM Ki16425 or 1 µM HA130 or 800 µg/mL heparin for 24 h. Ki16425, HA130, and heparin were added to cell culture 30 min prior to LPA, lysoPS, and PLA1A. The amounts of IL-8 released in the supernatants were monitored using ELISA. IL-8 produced by FLSs stimulated with LPA, lysoPS, and PLA1A was 18.17 ± 7.19, 55.65 ± 22.6, and 23.11 ± 7.44 pg/mL, respectively. For each experiment, stimulated IL-8 production without inhibitors was set at 100% for data normalization. The data are the means ± SE from three experiments. For statistical comparative analyses, cytokine amounts in cells stimulated with LPA or lysoPS or PLA1A were compared to that in cells stimulated in the presence of Ki16425 or HA130 or heparin. ** p < 0.01; *** p < 0.001.

Article Snippet: Normal human primary FLSs were purchased from Asterand (Detroit, MI, USA) or provided by Dr. Di Battista (MUHC-Research Institute).

Techniques: Expressing, Quantitative RT-PCR, Positive Control, Cell Culture, Enzyme-linked Immunosorbent Assay, Produced

Journal: International Journal of Molecular Sciences

Article Title: Phospholipase A1 Member A Activates Fibroblast-like Synoviocytes through the Autotaxin-Lysophosphatidic Acid Receptor Axis

doi: 10.3390/ijms222312685

Figure Lengend Snippet: IL-8 production in human primary FLSs stimulated with exogenous PLA1A and ATX. Human primary FLSs from normal donors were stimulated with 0.2 µg/mL recombinant PLA1A in combination with 5 nM recombinant ATX for 24 h, in the absence ( A ) or presence ( B ) of 1% fatty acid-free BSA added in the culture medium. Amounts of IL-8 released in the supernatants were monitored using ELISA. IL-8 produced by FLSs cultured in serum-free DMEM supplemented with or without 1% fatty acid-free BSA was 7.08 ± 2.45 and 8.05 ± 2.04 pg/mL, respectively. For each experiment, basal IL-8 production was set at 100% for data normalization. The data are the means ± SE from five ( A ) and three ( B ) experiments, respectively. For statistical comparative analyses, cytokine amounts in cells stimulated with PLA1A and/or ATX were compared to that in non-stimulated cells. ** p < 0.01.

Article Snippet: Normal human primary FLSs were purchased from Asterand (Detroit, MI, USA) or provided by Dr. Di Battista (MUHC-Research Institute).

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Produced, Cell Culture

Journal: The Plant Genome

Article Title: Combined transcriptomic and metabolomic analyses reveal the pharmacognostic mechanism of the metabolism of flavonoids in different parts of Polygonum capitatum

doi: 10.1002/tpg2.20543

Figure Lengend Snippet: Statistical bubble diagram of the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment of differentially expressed genes (DEGs). The abscissa represents the enrichment score, whereas the longitudinal axis represents the pathway term. The size of the bubbles corresponds to the number of enriched unigenes. The color of the bubble changes from blue to white to red. The smaller the enrichment p value is, the greater the degree of enrichment. (a) represents pcl versus pcf. (b) represents pcs versus pcf. (c) represents pcs versus pcl.

Article Snippet: Our transcriptomic results revealed that the expression levels of four unigenes encoding FLSs and one unigene encoding LAR were greater in P. capitatum flowers than in stems.

Techniques:

Journal: The Plant Genome

Article Title: Combined transcriptomic and metabolomic analyses reveal the pharmacognostic mechanism of the metabolism of flavonoids in different parts of Polygonum capitatum

doi: 10.1002/tpg2.20543

Figure Lengend Snippet: Heatmap diagram of differentially expressed genes (DEGs) related to flavonoid biosynthesis. The abscissa represents the number of samples, whereas the longitudinal axis represents the names of the unigenes. Red indicates upregulation, and light blue indicates downregulation. (a) represents pcl versus pcf. (b) represents pcs versus pcf. (c) represents pcs versus pcl.

Article Snippet: Our transcriptomic results revealed that the expression levels of four unigenes encoding FLSs and one unigene encoding LAR were greater in P. capitatum flowers than in stems.

Techniques:

Journal: The Plant Genome

Article Title: Combined transcriptomic and metabolomic analyses reveal the pharmacognostic mechanism of the metabolism of flavonoids in different parts of Polygonum capitatum

doi: 10.1002/tpg2.20543

Figure Lengend Snippet: Heatmap diagram of differentially expressed genes (DEGs) related to shikimic acid biosynthesis in different parts of Polygonum capitatum . The abscissa represents the number of samples, whereas the longitudinal axis represents the names of the unigenes. Red indicates upregulation, and light blue indicates downregulation. (a) represents pcl versus pcf. (b) represents pcs versus pcf. (c) represents pcs versus pcl.

Article Snippet: Our transcriptomic results revealed that the expression levels of four unigenes encoding FLSs and one unigene encoding LAR were greater in P. capitatum flowers than in stems.

Techniques: