Journal: Cells

Article Title: Dopamine Receptor Activation Modulates the Integrity of the Perisynaptic Extracellular Matrix at Excitatory Synapses

doi: 10.3390/cells9020260

Figure Lengend Snippet: siRNA sequences.

Article Snippet: The following primary antibodies were used: rabbit anti-ADAMTS-4 (Abcam, Cambridge, UK), rabbit anti-ADAMTS-5 (OriGene, Rockland, MD, USA), rabbit anti-aggrecan (Merck Millipore, Burlington, MA, USA), rabbit anti-aggrecan neoepitope (Novus Biologicals, Centennial, CO, USA), guinea pig anti-brevican (Seidenbecher et al., 1995) (custom-made; central region of rat BC), mouse anti-brevican (BD Biosciences, San José, CA, USA), rabbit anti-brevican “neo” (Rb399) (custom-made; neo-epitope CGGQEAVESE) [ , ], affinity-purified rabbit anti-brevican “neo” (Rb399) (custom-made; neo-epitope CGGQEAVESE), rat anti-D1 dopamine receptor (Sigma-Aldrich, St. Louis, MO, USA), rabbit anti-D2 dopamine receptor (Abcam, Cambridge, UK), mouse anti-GAD65 (Abcam, Cambridge, UK), rabbit anti-GAPDH (SYSY, Göttingen, Germany), rabbit anti-GFAP (SYSY, Göttingen, Germany), mouse anti-Homer 1 (SYSY, Göttingen, Germany), mouse anti-MAP2 (Sigma-Aldrich, St. Louis, MO, USA), and mouse anti-PSD95 (NeuroMab, Davis, CA, USA).

Techniques: shRNA

Journal: Cells

Article Title: Dopamine Receptor Activation Modulates the Integrity of the Perisynaptic Extracellular Matrix at Excitatory Synapses

doi: 10.3390/cells9020260

Figure Lengend Snippet: ADAMTS-4 and ADAMTS-5 are essential for D1-like-DA-receptor-induced BC cleavage. ( A ) Inhibition of ADAMTS-4 and ADAMTS-5 with TIMP3 resulted in a decrease in BC cleavage around excitatory synapses (Ctl, 1 ± 0.1013, n = 7; SKF, 1.623 ± 0.2566, n = 6; TIMP3, 0.7602 ± 0.0851, n = 6; SKF + TIMP3, 0.5882 ± 0.1017, n = 7; average ± SEM; one-way ANOVA; p = 0.0489; Dunnett’s multiple comparison test; * p < 0.05). ( B ) Knockdown of ADAMTS-4, ADAMTS-5, or both proteases together led to a significant decrease in D1-like-DA-receptor-induced BC cleavage (Ctl, 1 ± 0.0929, n = 6; SKF, 1.732 ± 0.134, n = 4; Scr, 1.091 ± 0.2033, n = 6; Scr + SKF, 1.773 ± 0.1169, n = 4; shA4, 0.3385 ± 0.1052, n = 4; shA4 + SKF, 0.5532 ± 0.1343, n = 5; shA5, 0.2658 ± 0.0362, n = 4; shA5 + SKF, 0.4606 ± 0.1183, n = 4; shA4 + shA5, 0.4122 ± 0.0619, n = 5; shA4 + shA5 + SKF, 0.4680 ± 0.0911, n = 6; average ± SEM; one-way ANOVA; P < 0.001; Dunnett’s multiple comparison test; *** p < 0.001) (* = significance compared to Ctl; # = significance compared to Scr). ( C ) ECM-modifying proteases were expressed in an inactive form carrying a pro-domain. Inhibition of the pro-protein convertase PACE4 diminished SKF81927-induced BC cleavage at excitatory synapses (Ctl, 1 ± 0.051, n = 6; SKF, 1.509 ± 0.1183, n = 6; Hexa-D-Arg, 0.9441 ± 0.0636, n = 6; Hexa-D-Arg + SKF, 0.8651 ± 0.1077, n = 4; average ± SEM; one-way ANOVA; p = 0.0002; Dunnett’s multiple comparison test; *** p < 0.001). (nFI cl. BC = normalized fluorescent intensity of cleaved brevican).

Article Snippet: The following primary antibodies were used: rabbit anti-ADAMTS-4 (Abcam, Cambridge, UK), rabbit anti-ADAMTS-5 (OriGene, Rockland, MD, USA), rabbit anti-aggrecan (Merck Millipore, Burlington, MA, USA), rabbit anti-aggrecan neoepitope (Novus Biologicals, Centennial, CO, USA), guinea pig anti-brevican (Seidenbecher et al., 1995) (custom-made; central region of rat BC), mouse anti-brevican (BD Biosciences, San José, CA, USA), rabbit anti-brevican “neo” (Rb399) (custom-made; neo-epitope CGGQEAVESE) [ , ], affinity-purified rabbit anti-brevican “neo” (Rb399) (custom-made; neo-epitope CGGQEAVESE), rat anti-D1 dopamine receptor (Sigma-Aldrich, St. Louis, MO, USA), rabbit anti-D2 dopamine receptor (Abcam, Cambridge, UK), mouse anti-GAD65 (Abcam, Cambridge, UK), rabbit anti-GAPDH (SYSY, Göttingen, Germany), rabbit anti-GFAP (SYSY, Göttingen, Germany), mouse anti-Homer 1 (SYSY, Göttingen, Germany), mouse anti-MAP2 (Sigma-Aldrich, St. Louis, MO, USA), and mouse anti-PSD95 (NeuroMab, Davis, CA, USA).

Techniques: Inhibition

Journal: Antibiotics

Article Title: Pathogenicity Islands in Uropathogenic Escherichia coli Clinical Isolate of the Globally Disseminated O25:H4-ST131 Pandemic Clonal Lineage: First Report from Egypt

doi: 10.3390/antibiotics11111620

Figure Lengend Snippet: ( A ) Circular genome comparison of EGY_EC14142 chromosome with other chromosomes of UPEC strains. Each of the genome sequence assemblies of UPEC strains was aligned against EGY_EC14142 using the Basic Local Alignment Search Tool nucleotide tool (BLASTn) and visualized using the BLAST Ring Image Generator (BRIG) tool. The outermost ring (red) corresponds to EGY_EC14142 chromosome belonging to EC14142 strain, with its size being displayed in the middle of the ring. Next, UTI89 (NC_007946) is shown (light green). The third ring corresponds to UMNO26 (NC_011751) (pink), then, 536 (NC_008253) (purple) and CFT073 (NC_004431) (dark green) are shown. Genomic regions covered by BLASTn are represented by a solid color in concentric rings (with varying color degrees depending on percentage identity), whereas white gaps indicate genomic regions not covered by BLASTn. ( B ) Phylogenetic tree of EC14142 relative to 22 UPEC strains. The phylogenetic tree was generated using MEGA software (v11.0.9) with the maximum likelihood approach according to the aligned sequences by Clustal Omega. E. coli K-12 MG1655 (NC_000913.3) was set as a reference, and it is displayed in black. Referral strains, 536 (NC_008253), CFT073 (NC_004431), UTI89 (NC_007946), and UMNO26 (NC_011751) are indicated by red bullets. A bootstrap replication value of 100 was applied.

Article Snippet: The BLAST Ring Image Generator (BRIG) tool ( http://sourceforge.net/projects/brig , accessed on 20 April 2022) was used to create a circular schematic map to compare the chromosome EGY_EC14142 to the chromosomes of other UPEC strains, namely, 536 (NC_008253), CFT073 (NC_004431), UTI89 (NC_007946), and UMN026 (NC_011751).

Techniques: Comparison, Sequencing, Generated, Software