Journal: Frontiers in pharmacology

Article Title: Dipyridamole activates adenosine A2B receptor and AMPK/cAMP signaling and promotes myogenic differentiation of myoblastic C2C12 cells.

doi: 10.3389/fphar.2023.1247664

Figure Lengend Snippet: FIGURE 3 Western blot protein expression over the 4 days of differentiation in the presence of tenofovir and dipyridamole. Protein expression of the A1, A2A, A2B, and A3 adenosine receptors, P2X7 receptor and Panx-1 is shown. Data are representative of n = 5 each group per day (mean ± SEM). *: treatment vs. basal; $: tenofovir and dipyridamole vs. control; #: dipyridamole and T + D vs. tenofovir. T + D: tenofovir + dipyridamole; DD: days of differentiation.

Article Snippet: Membranes were incubated overnight at 4°C with the primary antibodies for adenosine A1 (#55026-1- AP), Panx-1 (#12595-1-AP), CD39/ENTPD1 (#19229) (1:500), NT5E/CD73 (#12231) (1:1,000) (Proteintech, Rosemont, United States), A2A (# BS-1456R), A2B (#PA5-72850), A3 (#PA5-33326), MHC (#MA5-32555), PKAγ (#A25933) (Thermo Fisher Scientific, Waltham, United States), P2X7 (#ab109054), PKAβ (#ab187515), Mif5 (1:200) (#ab125078), MyoD (1:200) (#ab125078), myogenin (1:200) (#ab124800) (Abcam, Cambridge, United Kingdom), AMPK (#2532), pAMPK (Thr172) (#2535), PKAα (#4782), pCREB (Ser133) (#ab32096), CREB (#ab32515) (Cell signaling, Danvers, EEUU) (1:1,000 each) and Pax-7 (#AB_528428) (Developmental Studies Hybridoma Bank, Houston, EEUU) (1:500).

Techniques: Western Blot, Expressing, Control

Journal: Archives of toxicology

Article Title: Genetic ablation of pannexin1 counteracts liver fibrosis in a chemical, but not in a surgical mouse model

doi: 10.1007/s00204-018-2255-3

Figure Lengend Snippet: The Taqman probes and primers used for RT-qPCR analysis. Assay identification (ID) and accession number of target and reference genes are indicated. ( Actb , β-actin; ASC, apoptosis-associated speck-like protein containing a C-terminal caspase-recruitment domain; B2m , β-2-microglobulin; Casp1 , caspase 1; Clec7a , dectin-1; Col1a1 , collagen type 1 alpha 1; Cxcl14 , chemokine ligand 14; Gapdh , glyceraldehyde 3-phosphate dehydrogenase; Hmbs , hydroxymethylbilane synthase; Lbp , lipopolyssacharide-binding protein; Loxl2 , lysyl oxidase-like 2; Ly96 , lymphocyte antigen 96; Panx1 , pannexin1; S100a9 , S100 calcium-binding protein A9; Nalp3 , NACHT, LRR, and pyrin domain-containing protein 3; Ubc , ubiquitin C).

Article Snippet: Relative alterations (fold change) in RNA levels were calculated according to the 2 (-ΔΔCq) formula ( Livak and Schmittgen 2001 ). table ft1 table-wrap mode="anchored" t5 caption a7 Gene Assay ID Accession number Assay location Amplicon size (base pairs) Exon boundary Actb Mm00607939_s1 {"type":"entrez-nucleotide","attrs":{"text":"NM_007393.3","term_id":"145966868","term_text":"NM_007393.3"}} NM_007393.3 1233 115 6–6 ASC Mm00445747_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_023258.4","term_id":"165377184","term_text":"NM_023258.4"}} NM_023258.4 487 73 1-2 B2m Mm00437762_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_009735.3","term_id":"144227219","term_text":"NM_009735.3"}} NM_009735.3 111 77 1-2 Casp1 Mm00438023_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_009807.2","term_id":"86198304","term_text":"NM_009807.2"}} NM_009807.2 532 99 3-4 Col1a1 Mm00801666_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_007742.3","term_id":"118131144","term_text":"NM_007742.3"}} NM_007742.3 4071 89 49-50 Gapdh Mm99999915_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_008084.2","term_id":"126012538","term_text":"NM_008084.2"}} NM_008084.2 265 107 2–3 Hmbs Mm01143545_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_013551.2","term_id":"159110430","term_text":"NM_013551.2"}} NM_013551.2 473 81 6–7 Loxl2 Mm00804740_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_033325.2","term_id":"225579027","term_text":"NM_033325.2"}} NM_033325.2 2390 82 13-14 Nalp3 Mm00840904_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_145827.3","term_id":"118130221","term_text":"NM_145827.3"}} NM_145827.3 499 84 2-3 Panx1 Mm00450900_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_019482.2","term_id":"86262133","term_text":"NM_019482.2"}} NM_019482.2 986 70 3-4 Ubc Mm02525934_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_019639.4","term_id":"157671922","term_text":"NM_019639.4"}} NM_019639.4 370 176 2-2 Open in a separate window caption a8 The Taqman probes and primers used for RT-qPCR analysis.

Techniques: Ubiquitin Proteomics, Amplification

Journal: Archives of toxicology

Article Title: Genetic ablation of pannexin1 counteracts liver fibrosis in a chemical, but not in a surgical mouse model

doi: 10.1007/s00204-018-2255-3

Figure Lengend Snippet: WT and Panx1-/- mice (n = 5-15) were administered 20% CCl4 ip at a gradually increased dose for 8 weeks. (a) Area of fibrosis, area of α-SMA-positive cells, and Col1a1 and Loxl2 RNA levels were evaluated. (b) Serum levels of ALT, AST, albumin, total and conjugated bilirubin were determined. Results were analyzed by 1-way ANOVA followed by post hoc tests using Bonferroni correction. Data were expressed as means ± SEM (*p≤0.05; **p≤0.01; ***p≤0.001; ****p≤0.0001).

Article Snippet: Relative alterations (fold change) in RNA levels were calculated according to the 2 (-ΔΔCq) formula ( Livak and Schmittgen 2001 ). table ft1 table-wrap mode="anchored" t5 caption a7 Gene Assay ID Accession number Assay location Amplicon size (base pairs) Exon boundary Actb Mm00607939_s1 {"type":"entrez-nucleotide","attrs":{"text":"NM_007393.3","term_id":"145966868","term_text":"NM_007393.3"}} NM_007393.3 1233 115 6–6 ASC Mm00445747_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_023258.4","term_id":"165377184","term_text":"NM_023258.4"}} NM_023258.4 487 73 1-2 B2m Mm00437762_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_009735.3","term_id":"144227219","term_text":"NM_009735.3"}} NM_009735.3 111 77 1-2 Casp1 Mm00438023_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_009807.2","term_id":"86198304","term_text":"NM_009807.2"}} NM_009807.2 532 99 3-4 Col1a1 Mm00801666_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_007742.3","term_id":"118131144","term_text":"NM_007742.3"}} NM_007742.3 4071 89 49-50 Gapdh Mm99999915_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_008084.2","term_id":"126012538","term_text":"NM_008084.2"}} NM_008084.2 265 107 2–3 Hmbs Mm01143545_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_013551.2","term_id":"159110430","term_text":"NM_013551.2"}} NM_013551.2 473 81 6–7 Loxl2 Mm00804740_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_033325.2","term_id":"225579027","term_text":"NM_033325.2"}} NM_033325.2 2390 82 13-14 Nalp3 Mm00840904_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_145827.3","term_id":"118130221","term_text":"NM_145827.3"}} NM_145827.3 499 84 2-3 Panx1 Mm00450900_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_019482.2","term_id":"86262133","term_text":"NM_019482.2"}} NM_019482.2 986 70 3-4 Ubc Mm02525934_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_019639.4","term_id":"157671922","term_text":"NM_019639.4"}} NM_019639.4 370 176 2-2 Open in a separate window caption a8 The Taqman probes and primers used for RT-qPCR analysis.

Techniques:

Journal: Archives of toxicology

Article Title: Genetic ablation of pannexin1 counteracts liver fibrosis in a chemical, but not in a surgical mouse model

doi: 10.1007/s00204-018-2255-3

Figure Lengend Snippet: WT and Panx1-/- mice (n = 5-15) were subjected to BDL for 20 days. (a) Area of fibrosis, area of α-SMA-positive cells, and Col1a1 and Loxl2 RNA levels were evaluated. (b) Serum levels of ALP, ALT, AST, albumin, total and conjugated bilirubin were determined. Results were analyzed by 1-way ANOVA followed by post hoc tests using Bonferroni correction. Data were expressed as means ± SEM (*p≤0.05; ***p≤0.001; ****p≤0.0001).

Article Snippet: Relative alterations (fold change) in RNA levels were calculated according to the 2 (-ΔΔCq) formula ( Livak and Schmittgen 2001 ). table ft1 table-wrap mode="anchored" t5 caption a7 Gene Assay ID Accession number Assay location Amplicon size (base pairs) Exon boundary Actb Mm00607939_s1 {"type":"entrez-nucleotide","attrs":{"text":"NM_007393.3","term_id":"145966868","term_text":"NM_007393.3"}} NM_007393.3 1233 115 6–6 ASC Mm00445747_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_023258.4","term_id":"165377184","term_text":"NM_023258.4"}} NM_023258.4 487 73 1-2 B2m Mm00437762_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_009735.3","term_id":"144227219","term_text":"NM_009735.3"}} NM_009735.3 111 77 1-2 Casp1 Mm00438023_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_009807.2","term_id":"86198304","term_text":"NM_009807.2"}} NM_009807.2 532 99 3-4 Col1a1 Mm00801666_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_007742.3","term_id":"118131144","term_text":"NM_007742.3"}} NM_007742.3 4071 89 49-50 Gapdh Mm99999915_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_008084.2","term_id":"126012538","term_text":"NM_008084.2"}} NM_008084.2 265 107 2–3 Hmbs Mm01143545_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_013551.2","term_id":"159110430","term_text":"NM_013551.2"}} NM_013551.2 473 81 6–7 Loxl2 Mm00804740_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_033325.2","term_id":"225579027","term_text":"NM_033325.2"}} NM_033325.2 2390 82 13-14 Nalp3 Mm00840904_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_145827.3","term_id":"118130221","term_text":"NM_145827.3"}} NM_145827.3 499 84 2-3 Panx1 Mm00450900_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_019482.2","term_id":"86262133","term_text":"NM_019482.2"}} NM_019482.2 986 70 3-4 Ubc Mm02525934_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_019639.4","term_id":"157671922","term_text":"NM_019639.4"}} NM_019639.4 370 176 2-2 Open in a separate window caption a8 The Taqman probes and primers used for RT-qPCR analysis.

Techniques:

Journal: Archives of toxicology

Article Title: Genetic ablation of pannexin1 counteracts liver fibrosis in a chemical, but not in a surgical mouse model

doi: 10.1007/s00204-018-2255-3

Figure Lengend Snippet: WT and Panx1-/- mice (n = 8) were administered 20% CCl4 ip at a gradually increased dose for 8 weeks or subjected to BDL for 20 days. Activity of catalase, GPx, GR and SOD was evaluated in (a) the CCl4 model and (b) the BDL model. Results were analyzed by 1-way ANOVA followed by post hoc tests using Bonferroni correction and with Kruskal-Wallis test followed by post hoc tests using Dunn’s correction in case of SOD in the CCl4 model as well as catalase and SOD in the BDL model. Data were expressed as means ± SEM or median ± interquartile (*p≤0.05; **p≤0.01).

Article Snippet: Relative alterations (fold change) in RNA levels were calculated according to the 2 (-ΔΔCq) formula ( Livak and Schmittgen 2001 ). table ft1 table-wrap mode="anchored" t5 caption a7 Gene Assay ID Accession number Assay location Amplicon size (base pairs) Exon boundary Actb Mm00607939_s1 {"type":"entrez-nucleotide","attrs":{"text":"NM_007393.3","term_id":"145966868","term_text":"NM_007393.3"}} NM_007393.3 1233 115 6–6 ASC Mm00445747_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_023258.4","term_id":"165377184","term_text":"NM_023258.4"}} NM_023258.4 487 73 1-2 B2m Mm00437762_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_009735.3","term_id":"144227219","term_text":"NM_009735.3"}} NM_009735.3 111 77 1-2 Casp1 Mm00438023_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_009807.2","term_id":"86198304","term_text":"NM_009807.2"}} NM_009807.2 532 99 3-4 Col1a1 Mm00801666_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_007742.3","term_id":"118131144","term_text":"NM_007742.3"}} NM_007742.3 4071 89 49-50 Gapdh Mm99999915_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_008084.2","term_id":"126012538","term_text":"NM_008084.2"}} NM_008084.2 265 107 2–3 Hmbs Mm01143545_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_013551.2","term_id":"159110430","term_text":"NM_013551.2"}} NM_013551.2 473 81 6–7 Loxl2 Mm00804740_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_033325.2","term_id":"225579027","term_text":"NM_033325.2"}} NM_033325.2 2390 82 13-14 Nalp3 Mm00840904_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_145827.3","term_id":"118130221","term_text":"NM_145827.3"}} NM_145827.3 499 84 2-3 Panx1 Mm00450900_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_019482.2","term_id":"86262133","term_text":"NM_019482.2"}} NM_019482.2 986 70 3-4 Ubc Mm02525934_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_019639.4","term_id":"157671922","term_text":"NM_019639.4"}} NM_019639.4 370 176 2-2 Open in a separate window caption a8 The Taqman probes and primers used for RT-qPCR analysis.

Techniques: Activity Assay

Journal: Archives of toxicology

Article Title: Genetic ablation of pannexin1 counteracts liver fibrosis in a chemical, but not in a surgical mouse model

doi: 10.1007/s00204-018-2255-3

Figure Lengend Snippet: WT and Panx1-/- mice were administered 20% CCl4 ip at a gradually increased dose for 8 weeks or subjected to BDL for 20 days. Representative images for the calculation of hepatic CD68+ cell density (n = 5) and example of blot membranes for the determination of p-NF-κBp65 S536 protein levels (n = 9 – 15) in (a) the CCl4 model and (b) the BDL model. Results were analyzed by 1-way ANOVA followed by post hoc tests using Bonferroni correction and with Kruskal-Wallis test followed by post hoc tests using Dunn’s correction in case of the p-NF-κBp65 S536. Data were expressed as means ± SEM or median ± interquartile (*p≤0.05; ***p≤0.001; **** p≤0.0001). Scale bar represents 20 μm.

Article Snippet: Relative alterations (fold change) in RNA levels were calculated according to the 2 (-ΔΔCq) formula ( Livak and Schmittgen 2001 ). table ft1 table-wrap mode="anchored" t5 caption a7 Gene Assay ID Accession number Assay location Amplicon size (base pairs) Exon boundary Actb Mm00607939_s1 {"type":"entrez-nucleotide","attrs":{"text":"NM_007393.3","term_id":"145966868","term_text":"NM_007393.3"}} NM_007393.3 1233 115 6–6 ASC Mm00445747_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_023258.4","term_id":"165377184","term_text":"NM_023258.4"}} NM_023258.4 487 73 1-2 B2m Mm00437762_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_009735.3","term_id":"144227219","term_text":"NM_009735.3"}} NM_009735.3 111 77 1-2 Casp1 Mm00438023_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_009807.2","term_id":"86198304","term_text":"NM_009807.2"}} NM_009807.2 532 99 3-4 Col1a1 Mm00801666_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_007742.3","term_id":"118131144","term_text":"NM_007742.3"}} NM_007742.3 4071 89 49-50 Gapdh Mm99999915_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_008084.2","term_id":"126012538","term_text":"NM_008084.2"}} NM_008084.2 265 107 2–3 Hmbs Mm01143545_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_013551.2","term_id":"159110430","term_text":"NM_013551.2"}} NM_013551.2 473 81 6–7 Loxl2 Mm00804740_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_033325.2","term_id":"225579027","term_text":"NM_033325.2"}} NM_033325.2 2390 82 13-14 Nalp3 Mm00840904_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_145827.3","term_id":"118130221","term_text":"NM_145827.3"}} NM_145827.3 499 84 2-3 Panx1 Mm00450900_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_019482.2","term_id":"86262133","term_text":"NM_019482.2"}} NM_019482.2 986 70 3-4 Ubc Mm02525934_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_019639.4","term_id":"157671922","term_text":"NM_019639.4"}} NM_019639.4 370 176 2-2 Open in a separate window caption a8 The Taqman probes and primers used for RT-qPCR analysis.

Techniques:

Journal: Archives of toxicology

Article Title: Genetic ablation of pannexin1 counteracts liver fibrosis in a chemical, but not in a surgical mouse model

doi: 10.1007/s00204-018-2255-3

Figure Lengend Snippet: WT and Panx1-/- mice (n = 10) were administered 20% CCl4 ip at a gradually increased dose for 8 weeks. Results were analyzed by 1-way ANOVA followed by post hoc tests using Bonferroni correction. Data were expressed as means ± SEM (***p≤0.001; ****p≤0.0001).

Article Snippet: Relative alterations (fold change) in RNA levels were calculated according to the 2 (-ΔΔCq) formula ( Livak and Schmittgen 2001 ). table ft1 table-wrap mode="anchored" t5 caption a7 Gene Assay ID Accession number Assay location Amplicon size (base pairs) Exon boundary Actb Mm00607939_s1 {"type":"entrez-nucleotide","attrs":{"text":"NM_007393.3","term_id":"145966868","term_text":"NM_007393.3"}} NM_007393.3 1233 115 6–6 ASC Mm00445747_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_023258.4","term_id":"165377184","term_text":"NM_023258.4"}} NM_023258.4 487 73 1-2 B2m Mm00437762_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_009735.3","term_id":"144227219","term_text":"NM_009735.3"}} NM_009735.3 111 77 1-2 Casp1 Mm00438023_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_009807.2","term_id":"86198304","term_text":"NM_009807.2"}} NM_009807.2 532 99 3-4 Col1a1 Mm00801666_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_007742.3","term_id":"118131144","term_text":"NM_007742.3"}} NM_007742.3 4071 89 49-50 Gapdh Mm99999915_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_008084.2","term_id":"126012538","term_text":"NM_008084.2"}} NM_008084.2 265 107 2–3 Hmbs Mm01143545_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_013551.2","term_id":"159110430","term_text":"NM_013551.2"}} NM_013551.2 473 81 6–7 Loxl2 Mm00804740_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_033325.2","term_id":"225579027","term_text":"NM_033325.2"}} NM_033325.2 2390 82 13-14 Nalp3 Mm00840904_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_145827.3","term_id":"118130221","term_text":"NM_145827.3"}} NM_145827.3 499 84 2-3 Panx1 Mm00450900_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_019482.2","term_id":"86262133","term_text":"NM_019482.2"}} NM_019482.2 986 70 3-4 Ubc Mm02525934_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_019639.4","term_id":"157671922","term_text":"NM_019639.4"}} NM_019639.4 370 176 2-2 Open in a separate window caption a8 The Taqman probes and primers used for RT-qPCR analysis.

Techniques:

Journal: Archives of toxicology

Article Title: Genetic ablation of pannexin1 counteracts liver fibrosis in a chemical, but not in a surgical mouse model

doi: 10.1007/s00204-018-2255-3

Figure Lengend Snippet: WT and Panx1-/- mice (n = 4) were administered 20% CCl4 ip at a gradually increased dose for 8 weeks. Representative antibody array membranes from liver tissue extract corresponded to WT and Panx1-/- treated with oil or CCl4. Data were analyzed by densitometry analysis. Then, background was subtracted and results were normalized against the average of positive control. Results were analyzed by 1-way ANOVA followed by post hoc tests using Bonferroni correction. Data were expressed as means ± SEM (*p≤0.05; **p≤0.01; ***p≤0.001).

Article Snippet: Relative alterations (fold change) in RNA levels were calculated according to the 2 (-ΔΔCq) formula ( Livak and Schmittgen 2001 ). table ft1 table-wrap mode="anchored" t5 caption a7 Gene Assay ID Accession number Assay location Amplicon size (base pairs) Exon boundary Actb Mm00607939_s1 {"type":"entrez-nucleotide","attrs":{"text":"NM_007393.3","term_id":"145966868","term_text":"NM_007393.3"}} NM_007393.3 1233 115 6–6 ASC Mm00445747_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_023258.4","term_id":"165377184","term_text":"NM_023258.4"}} NM_023258.4 487 73 1-2 B2m Mm00437762_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_009735.3","term_id":"144227219","term_text":"NM_009735.3"}} NM_009735.3 111 77 1-2 Casp1 Mm00438023_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_009807.2","term_id":"86198304","term_text":"NM_009807.2"}} NM_009807.2 532 99 3-4 Col1a1 Mm00801666_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_007742.3","term_id":"118131144","term_text":"NM_007742.3"}} NM_007742.3 4071 89 49-50 Gapdh Mm99999915_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_008084.2","term_id":"126012538","term_text":"NM_008084.2"}} NM_008084.2 265 107 2–3 Hmbs Mm01143545_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_013551.2","term_id":"159110430","term_text":"NM_013551.2"}} NM_013551.2 473 81 6–7 Loxl2 Mm00804740_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_033325.2","term_id":"225579027","term_text":"NM_033325.2"}} NM_033325.2 2390 82 13-14 Nalp3 Mm00840904_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_145827.3","term_id":"118130221","term_text":"NM_145827.3"}} NM_145827.3 499 84 2-3 Panx1 Mm00450900_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_019482.2","term_id":"86262133","term_text":"NM_019482.2"}} NM_019482.2 986 70 3-4 Ubc Mm02525934_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_019639.4","term_id":"157671922","term_text":"NM_019639.4"}} NM_019639.4 370 176 2-2 Open in a separate window caption a8 The Taqman probes and primers used for RT-qPCR analysis.

Techniques: Ab Array, Positive Control

Journal: Archives of toxicology

Article Title: Genetic ablation of pannexin1 counteracts liver fibrosis in a chemical, but not in a surgical mouse model

doi: 10.1007/s00204-018-2255-3

Figure Lengend Snippet: WT and Panx1-/- mice (n = 4) were subjected to BDL for 20 days. Representative antibody array membranes from liver tissue extract corresponded to WT and Panx1-/- subjected to sham or BDL. Data were analyzed by densitometry analysis. Then, background was substracted and results were normalized against the average of positive control. Results were analyzed by 1-way ANOVA followed by post hoc tests using Bonferroni correction. Data were expressed as means ± SEM (*p≤0.05; **p≤0.01; ***p≤0.001; ****p≤0.0001).

Article Snippet: Relative alterations (fold change) in RNA levels were calculated according to the 2 (-ΔΔCq) formula ( Livak and Schmittgen 2001 ). table ft1 table-wrap mode="anchored" t5 caption a7 Gene Assay ID Accession number Assay location Amplicon size (base pairs) Exon boundary Actb Mm00607939_s1 {"type":"entrez-nucleotide","attrs":{"text":"NM_007393.3","term_id":"145966868","term_text":"NM_007393.3"}} NM_007393.3 1233 115 6–6 ASC Mm00445747_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_023258.4","term_id":"165377184","term_text":"NM_023258.4"}} NM_023258.4 487 73 1-2 B2m Mm00437762_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_009735.3","term_id":"144227219","term_text":"NM_009735.3"}} NM_009735.3 111 77 1-2 Casp1 Mm00438023_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_009807.2","term_id":"86198304","term_text":"NM_009807.2"}} NM_009807.2 532 99 3-4 Col1a1 Mm00801666_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_007742.3","term_id":"118131144","term_text":"NM_007742.3"}} NM_007742.3 4071 89 49-50 Gapdh Mm99999915_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_008084.2","term_id":"126012538","term_text":"NM_008084.2"}} NM_008084.2 265 107 2–3 Hmbs Mm01143545_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_013551.2","term_id":"159110430","term_text":"NM_013551.2"}} NM_013551.2 473 81 6–7 Loxl2 Mm00804740_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_033325.2","term_id":"225579027","term_text":"NM_033325.2"}} NM_033325.2 2390 82 13-14 Nalp3 Mm00840904_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_145827.3","term_id":"118130221","term_text":"NM_145827.3"}} NM_145827.3 499 84 2-3 Panx1 Mm00450900_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_019482.2","term_id":"86262133","term_text":"NM_019482.2"}} NM_019482.2 986 70 3-4 Ubc Mm02525934_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_019639.4","term_id":"157671922","term_text":"NM_019639.4"}} NM_019639.4 370 176 2-2 Open in a separate window caption a8 The Taqman probes and primers used for RT-qPCR analysis.

Techniques: Ab Array, Positive Control

Journal: Archives of toxicology

Article Title: Genetic ablation of pannexin1 counteracts liver fibrosis in a chemical, but not in a surgical mouse model

doi: 10.1007/s00204-018-2255-3

Figure Lengend Snippet: WT and Panx1-/- mice (n = 3) were administered 20% CCl4 ip at a gradually increased dose for 8 weeks. Representative images from CD11b, Ly6c, Ly6g and CD3 immune markers.

Article Snippet: Relative alterations (fold change) in RNA levels were calculated according to the 2 (-ΔΔCq) formula ( Livak and Schmittgen 2001 ). table ft1 table-wrap mode="anchored" t5 caption a7 Gene Assay ID Accession number Assay location Amplicon size (base pairs) Exon boundary Actb Mm00607939_s1 {"type":"entrez-nucleotide","attrs":{"text":"NM_007393.3","term_id":"145966868","term_text":"NM_007393.3"}} NM_007393.3 1233 115 6–6 ASC Mm00445747_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_023258.4","term_id":"165377184","term_text":"NM_023258.4"}} NM_023258.4 487 73 1-2 B2m Mm00437762_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_009735.3","term_id":"144227219","term_text":"NM_009735.3"}} NM_009735.3 111 77 1-2 Casp1 Mm00438023_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_009807.2","term_id":"86198304","term_text":"NM_009807.2"}} NM_009807.2 532 99 3-4 Col1a1 Mm00801666_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_007742.3","term_id":"118131144","term_text":"NM_007742.3"}} NM_007742.3 4071 89 49-50 Gapdh Mm99999915_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_008084.2","term_id":"126012538","term_text":"NM_008084.2"}} NM_008084.2 265 107 2–3 Hmbs Mm01143545_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_013551.2","term_id":"159110430","term_text":"NM_013551.2"}} NM_013551.2 473 81 6–7 Loxl2 Mm00804740_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_033325.2","term_id":"225579027","term_text":"NM_033325.2"}} NM_033325.2 2390 82 13-14 Nalp3 Mm00840904_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_145827.3","term_id":"118130221","term_text":"NM_145827.3"}} NM_145827.3 499 84 2-3 Panx1 Mm00450900_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_019482.2","term_id":"86262133","term_text":"NM_019482.2"}} NM_019482.2 986 70 3-4 Ubc Mm02525934_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_019639.4","term_id":"157671922","term_text":"NM_019639.4"}} NM_019639.4 370 176 2-2 Open in a separate window caption a8 The Taqman probes and primers used for RT-qPCR analysis.

Techniques:

Journal: Archives of toxicology

Article Title: Genetic ablation of pannexin1 counteracts liver fibrosis in a chemical, but not in a surgical mouse model

doi: 10.1007/s00204-018-2255-3

Figure Lengend Snippet: WT and Panx1-/- mice (n = 3) were subjected to BDL for 20 days. Representative images from CD11b, Ly6c, Ly6g and CD3 immune markers.

Article Snippet: Relative alterations (fold change) in RNA levels were calculated according to the 2 (-ΔΔCq) formula ( Livak and Schmittgen 2001 ). table ft1 table-wrap mode="anchored" t5 caption a7 Gene Assay ID Accession number Assay location Amplicon size (base pairs) Exon boundary Actb Mm00607939_s1 {"type":"entrez-nucleotide","attrs":{"text":"NM_007393.3","term_id":"145966868","term_text":"NM_007393.3"}} NM_007393.3 1233 115 6–6 ASC Mm00445747_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_023258.4","term_id":"165377184","term_text":"NM_023258.4"}} NM_023258.4 487 73 1-2 B2m Mm00437762_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_009735.3","term_id":"144227219","term_text":"NM_009735.3"}} NM_009735.3 111 77 1-2 Casp1 Mm00438023_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_009807.2","term_id":"86198304","term_text":"NM_009807.2"}} NM_009807.2 532 99 3-4 Col1a1 Mm00801666_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_007742.3","term_id":"118131144","term_text":"NM_007742.3"}} NM_007742.3 4071 89 49-50 Gapdh Mm99999915_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_008084.2","term_id":"126012538","term_text":"NM_008084.2"}} NM_008084.2 265 107 2–3 Hmbs Mm01143545_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_013551.2","term_id":"159110430","term_text":"NM_013551.2"}} NM_013551.2 473 81 6–7 Loxl2 Mm00804740_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_033325.2","term_id":"225579027","term_text":"NM_033325.2"}} NM_033325.2 2390 82 13-14 Nalp3 Mm00840904_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_145827.3","term_id":"118130221","term_text":"NM_145827.3"}} NM_145827.3 499 84 2-3 Panx1 Mm00450900_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_019482.2","term_id":"86262133","term_text":"NM_019482.2"}} NM_019482.2 986 70 3-4 Ubc Mm02525934_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_019639.4","term_id":"157671922","term_text":"NM_019639.4"}} NM_019639.4 370 176 2-2 Open in a separate window caption a8 The Taqman probes and primers used for RT-qPCR analysis.

Techniques:

Journal: Archives of toxicology

Article Title: Genetic ablation of pannexin1 counteracts liver fibrosis in a chemical, but not in a surgical mouse model

doi: 10.1007/s00204-018-2255-3

Figure Lengend Snippet: WT and Panx1-/- mice (n = 5) were administered 20% CCl4 ip at a gradually increased dose for 8 weeks or subjected to BDL for 20 days. Ki-67+ cell density in (a) the CCl4 model and (b) the BDL model. Results were analyzed by 1-way ANOVA followed by post hoc tests using Bonferroni correction and with Kruskal-Wallis test followed by post hoc tests using Dunn’s correction in case of the Ki-67+ hepatocytes and cholangiocytes within the BDL model. Data were expressed as means ± SEM or median ± interquartile (*p≤0.05; **** p≤0.0001). Scale bar represents 20 μm.

Article Snippet: Relative alterations (fold change) in RNA levels were calculated according to the 2 (-ΔΔCq) formula ( Livak and Schmittgen 2001 ). table ft1 table-wrap mode="anchored" t5 caption a7 Gene Assay ID Accession number Assay location Amplicon size (base pairs) Exon boundary Actb Mm00607939_s1 {"type":"entrez-nucleotide","attrs":{"text":"NM_007393.3","term_id":"145966868","term_text":"NM_007393.3"}} NM_007393.3 1233 115 6–6 ASC Mm00445747_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_023258.4","term_id":"165377184","term_text":"NM_023258.4"}} NM_023258.4 487 73 1-2 B2m Mm00437762_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_009735.3","term_id":"144227219","term_text":"NM_009735.3"}} NM_009735.3 111 77 1-2 Casp1 Mm00438023_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_009807.2","term_id":"86198304","term_text":"NM_009807.2"}} NM_009807.2 532 99 3-4 Col1a1 Mm00801666_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_007742.3","term_id":"118131144","term_text":"NM_007742.3"}} NM_007742.3 4071 89 49-50 Gapdh Mm99999915_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_008084.2","term_id":"126012538","term_text":"NM_008084.2"}} NM_008084.2 265 107 2–3 Hmbs Mm01143545_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_013551.2","term_id":"159110430","term_text":"NM_013551.2"}} NM_013551.2 473 81 6–7 Loxl2 Mm00804740_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_033325.2","term_id":"225579027","term_text":"NM_033325.2"}} NM_033325.2 2390 82 13-14 Nalp3 Mm00840904_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_145827.3","term_id":"118130221","term_text":"NM_145827.3"}} NM_145827.3 499 84 2-3 Panx1 Mm00450900_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_019482.2","term_id":"86262133","term_text":"NM_019482.2"}} NM_019482.2 986 70 3-4 Ubc Mm02525934_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_019639.4","term_id":"157671922","term_text":"NM_019639.4"}} NM_019639.4 370 176 2-2 Open in a separate window caption a8 The Taqman probes and primers used for RT-qPCR analysis.

Techniques:

Journal: Archives of toxicology

Article Title: Genetic ablation of pannexin1 counteracts liver fibrosis in a chemical, but not in a surgical mouse model

doi: 10.1007/s00204-018-2255-3

Figure Lengend Snippet: Gene modulation by CCl 4 treatment in Panx1 -/- genetic background.

Article Snippet: Relative alterations (fold change) in RNA levels were calculated according to the 2 (-ΔΔCq) formula ( Livak and Schmittgen 2001 ). table ft1 table-wrap mode="anchored" t5 caption a7 Gene Assay ID Accession number Assay location Amplicon size (base pairs) Exon boundary Actb Mm00607939_s1 {"type":"entrez-nucleotide","attrs":{"text":"NM_007393.3","term_id":"145966868","term_text":"NM_007393.3"}} NM_007393.3 1233 115 6–6 ASC Mm00445747_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_023258.4","term_id":"165377184","term_text":"NM_023258.4"}} NM_023258.4 487 73 1-2 B2m Mm00437762_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_009735.3","term_id":"144227219","term_text":"NM_009735.3"}} NM_009735.3 111 77 1-2 Casp1 Mm00438023_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_009807.2","term_id":"86198304","term_text":"NM_009807.2"}} NM_009807.2 532 99 3-4 Col1a1 Mm00801666_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_007742.3","term_id":"118131144","term_text":"NM_007742.3"}} NM_007742.3 4071 89 49-50 Gapdh Mm99999915_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_008084.2","term_id":"126012538","term_text":"NM_008084.2"}} NM_008084.2 265 107 2–3 Hmbs Mm01143545_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_013551.2","term_id":"159110430","term_text":"NM_013551.2"}} NM_013551.2 473 81 6–7 Loxl2 Mm00804740_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_033325.2","term_id":"225579027","term_text":"NM_033325.2"}} NM_033325.2 2390 82 13-14 Nalp3 Mm00840904_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_145827.3","term_id":"118130221","term_text":"NM_145827.3"}} NM_145827.3 499 84 2-3 Panx1 Mm00450900_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_019482.2","term_id":"86262133","term_text":"NM_019482.2"}} NM_019482.2 986 70 3-4 Ubc Mm02525934_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_019639.4","term_id":"157671922","term_text":"NM_019639.4"}} NM_019639.4 370 176 2-2 Open in a separate window caption a8 The Taqman probes and primers used for RT-qPCR analysis.

Techniques: Functional Assay, Binding Assay, Activity Assay

Journal: Archives of toxicology

Article Title: Genetic ablation of pannexin1 counteracts liver fibrosis in a chemical, but not in a surgical mouse model

doi: 10.1007/s00204-018-2255-3

Figure Lengend Snippet: WT and Panx1-/- mice (n = 5) administered 20% CCl4 ip at a gradually increased dose for 8 weeks. (a) Gene ontogeny of the different clusters of modulated genes expressed as the average of the fold change from genes with at least 2-fold difference and p<0.05 and (b) RT-qPCR analysis from genes corresponded to the categories: CCl4 independent of genetic background or CCl4 in Panx1-/- genetic background. Results were analyzed by 1-way ANOVA followed by post hoc tests using Bonferroni correction. Data were expressed as means ± SEM (**p≤0.01; ****p≤0.0001). (N.M., not modulated).

Article Snippet: Relative alterations (fold change) in RNA levels were calculated according to the 2 (-ΔΔCq) formula ( Livak and Schmittgen 2001 ). table ft1 table-wrap mode="anchored" t5 caption a7 Gene Assay ID Accession number Assay location Amplicon size (base pairs) Exon boundary Actb Mm00607939_s1 {"type":"entrez-nucleotide","attrs":{"text":"NM_007393.3","term_id":"145966868","term_text":"NM_007393.3"}} NM_007393.3 1233 115 6–6 ASC Mm00445747_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_023258.4","term_id":"165377184","term_text":"NM_023258.4"}} NM_023258.4 487 73 1-2 B2m Mm00437762_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_009735.3","term_id":"144227219","term_text":"NM_009735.3"}} NM_009735.3 111 77 1-2 Casp1 Mm00438023_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_009807.2","term_id":"86198304","term_text":"NM_009807.2"}} NM_009807.2 532 99 3-4 Col1a1 Mm00801666_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_007742.3","term_id":"118131144","term_text":"NM_007742.3"}} NM_007742.3 4071 89 49-50 Gapdh Mm99999915_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_008084.2","term_id":"126012538","term_text":"NM_008084.2"}} NM_008084.2 265 107 2–3 Hmbs Mm01143545_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_013551.2","term_id":"159110430","term_text":"NM_013551.2"}} NM_013551.2 473 81 6–7 Loxl2 Mm00804740_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_033325.2","term_id":"225579027","term_text":"NM_033325.2"}} NM_033325.2 2390 82 13-14 Nalp3 Mm00840904_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_145827.3","term_id":"118130221","term_text":"NM_145827.3"}} NM_145827.3 499 84 2-3 Panx1 Mm00450900_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_019482.2","term_id":"86262133","term_text":"NM_019482.2"}} NM_019482.2 986 70 3-4 Ubc Mm02525934_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_019639.4","term_id":"157671922","term_text":"NM_019639.4"}} NM_019639.4 370 176 2-2 Open in a separate window caption a8 The Taqman probes and primers used for RT-qPCR analysis.

Techniques: Quantitative RT-PCR

Journal: Archives of toxicology

Article Title: Genetic ablation of pannexin1 counteracts liver fibrosis in a chemical, but not in a surgical mouse model

doi: 10.1007/s00204-018-2255-3

Figure Lengend Snippet: Gene modulation by the BDL procedure in Panx1 -/- genetic background.

Article Snippet: Relative alterations (fold change) in RNA levels were calculated according to the 2 (-ΔΔCq) formula ( Livak and Schmittgen 2001 ). table ft1 table-wrap mode="anchored" t5 caption a7 Gene Assay ID Accession number Assay location Amplicon size (base pairs) Exon boundary Actb Mm00607939_s1 {"type":"entrez-nucleotide","attrs":{"text":"NM_007393.3","term_id":"145966868","term_text":"NM_007393.3"}} NM_007393.3 1233 115 6–6 ASC Mm00445747_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_023258.4","term_id":"165377184","term_text":"NM_023258.4"}} NM_023258.4 487 73 1-2 B2m Mm00437762_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_009735.3","term_id":"144227219","term_text":"NM_009735.3"}} NM_009735.3 111 77 1-2 Casp1 Mm00438023_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_009807.2","term_id":"86198304","term_text":"NM_009807.2"}} NM_009807.2 532 99 3-4 Col1a1 Mm00801666_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_007742.3","term_id":"118131144","term_text":"NM_007742.3"}} NM_007742.3 4071 89 49-50 Gapdh Mm99999915_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_008084.2","term_id":"126012538","term_text":"NM_008084.2"}} NM_008084.2 265 107 2–3 Hmbs Mm01143545_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_013551.2","term_id":"159110430","term_text":"NM_013551.2"}} NM_013551.2 473 81 6–7 Loxl2 Mm00804740_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_033325.2","term_id":"225579027","term_text":"NM_033325.2"}} NM_033325.2 2390 82 13-14 Nalp3 Mm00840904_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_145827.3","term_id":"118130221","term_text":"NM_145827.3"}} NM_145827.3 499 84 2-3 Panx1 Mm00450900_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_019482.2","term_id":"86262133","term_text":"NM_019482.2"}} NM_019482.2 986 70 3-4 Ubc Mm02525934_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_019639.4","term_id":"157671922","term_text":"NM_019639.4"}} NM_019639.4 370 176 2-2 Open in a separate window caption a8 The Taqman probes and primers used for RT-qPCR analysis.

Techniques: Functional Assay, Binding Assay, Membrane, Activity Assay, Ubiquitin Proteomics

Journal: Archives of toxicology

Article Title: Genetic ablation of pannexin1 counteracts liver fibrosis in a chemical, but not in a surgical mouse model

doi: 10.1007/s00204-018-2255-3

Figure Lengend Snippet: WT and Panx1-/- mice (n = 5) subjected to BDL for 20 days. (a) Gene ontogeny of the different clusters of modulated genes expressed as the average of the fold change from genes with at least 2-fold difference and p<0.05 and (b) RT-qPCR analysis from genes corresponded to the categories: BDL independent of genetic background or BDL in Panx1-/- genetic background. Results were analyzed by 1-way ANOVA followed by post hoc tests using Bonferroni correction. Data were expressed as means ± SEM (***p≤0.001). (N.M., not modulated).

Article Snippet: Relative alterations (fold change) in RNA levels were calculated according to the 2 (-ΔΔCq) formula ( Livak and Schmittgen 2001 ). table ft1 table-wrap mode="anchored" t5 caption a7 Gene Assay ID Accession number Assay location Amplicon size (base pairs) Exon boundary Actb Mm00607939_s1 {"type":"entrez-nucleotide","attrs":{"text":"NM_007393.3","term_id":"145966868","term_text":"NM_007393.3"}} NM_007393.3 1233 115 6–6 ASC Mm00445747_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_023258.4","term_id":"165377184","term_text":"NM_023258.4"}} NM_023258.4 487 73 1-2 B2m Mm00437762_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_009735.3","term_id":"144227219","term_text":"NM_009735.3"}} NM_009735.3 111 77 1-2 Casp1 Mm00438023_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_009807.2","term_id":"86198304","term_text":"NM_009807.2"}} NM_009807.2 532 99 3-4 Col1a1 Mm00801666_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_007742.3","term_id":"118131144","term_text":"NM_007742.3"}} NM_007742.3 4071 89 49-50 Gapdh Mm99999915_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_008084.2","term_id":"126012538","term_text":"NM_008084.2"}} NM_008084.2 265 107 2–3 Hmbs Mm01143545_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_013551.2","term_id":"159110430","term_text":"NM_013551.2"}} NM_013551.2 473 81 6–7 Loxl2 Mm00804740_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_033325.2","term_id":"225579027","term_text":"NM_033325.2"}} NM_033325.2 2390 82 13-14 Nalp3 Mm00840904_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_145827.3","term_id":"118130221","term_text":"NM_145827.3"}} NM_145827.3 499 84 2-3 Panx1 Mm00450900_m1 {"type":"entrez-nucleotide","attrs":{"text":"NM_019482.2","term_id":"86262133","term_text":"NM_019482.2"}} NM_019482.2 986 70 3-4 Ubc Mm02525934_g1 {"type":"entrez-nucleotide","attrs":{"text":"NM_019639.4","term_id":"157671922","term_text":"NM_019639.4"}} NM_019639.4 370 176 2-2 Open in a separate window caption a8 The Taqman probes and primers used for RT-qPCR analysis.

Techniques: Quantitative RT-PCR

Journal: bioRxiv

Article Title: Pannexin 1 Channels Control Cardiomyocyte Metabolism and Neutrophil Recruitment During Non-Ischemic Heart Failure

doi: 10.1101/2023.12.29.573679

Figure Lengend Snippet: A) H9c2 were transfected with either control or Panx1-targeting siRNA for 72 hours and extracellular concentration of ATP was measured after 15 minutes of stimulation with isoproterenol (ISO) (2 and 20μM) or vehicle control. (N = 4) B) Flow cytometric analysis of Yo-Pro-1 dye uptake by control of Panx1-targeting siRNA treated H9c2 cells after stimulation with 20μM ISO (gated for live, annexin V - cells). (N = 4) C) H9c2 cells were pretreated with spironolactone (50μM) or vehicle control and extracellular ATP levels were measured after 15 minutes of stimulation with isoproterenol (ISO) (2 and 20μM). (N = 4) D) Yo-Pro-1 dye uptake by H9c2 cells treated with isoproterenol (20μM) after pretreatment with spironolactone, carbenoxolone (CBX), or probenecid at the listed concentrations. (N = 4) Data are represented as mean ± standard error of the mean (SEM). Data shown are representatives of 2 independent experiments. Significance determined using One-way ANOVA (A, B, C, D) with Sidak’s multiple comparison test for post-hoc analysis for comparisons between individual groups. ns = not significant See also S1 in reference to .

Article Snippet: Quantitative reverse transcription PCR was performed using TaqManFast Advanced Master Mix (Thermo Fisher Scientific) and TaqMan primers (mouse Panx1 mm0045091 – Invitrogen) (Rat Panx1 Rn01447976_m1 – Thermo Fisher Scientific) for genes of interest on a CFX Connect real-time PCR instrument (Bio-Rad).

Techniques: Transfection, Control, Concentration Assay, Comparison

Journal: bioRxiv

Article Title: Pannexin 1 Channels Control Cardiomyocyte Metabolism and Neutrophil Recruitment During Non-Ischemic Heart Failure

doi: 10.1101/2023.12.29.573679

Figure Lengend Snippet: A) Confocal micrographs of cardiomyocyte-rich whole heart sections from 12-week-old male Panx1 fl/fl and Panx1 MyHC6 mice. Knockout of PANX1 in cardiomyocytes was assessed using immunofluorescent staining. Cardiac troponin T (cTnnt) was used to stain cardiomyocytes; DAPI was used as a nuclear stain. Left, representative micrographs with insets showing PANX1 punctate; middle, average Panx1 fluorescence intensity for individual mice; right, fluorescence intensities for individual micrographs showing demonstrating of PANX1 signal. (Panx1 fl/fl N = 4 mice N = 13 images, Panx1 MyHC6 N = 3 mice N = 9 images, 3-5 images per mouse averaged). B) End-diastolic volume C) End-systolic volume D) Ejection fraction and E) Wall thickness were measured in anesthetized 8- to 10-week-old male Panx1 fl/fl and Panx1 MyHC6 mice using in vivo 2D-echocardiography. (Panx1 fl/fl N = 9 mice, Panx1 MyHC6 N = 8) Data represented as mean ± SEM. Significance was determined by Welch’s t-test (A-E). ns = not significant. See also S2 in reference to .

Article Snippet: Quantitative reverse transcription PCR was performed using TaqManFast Advanced Master Mix (Thermo Fisher Scientific) and TaqMan primers (mouse Panx1 mm0045091 – Invitrogen) (Rat Panx1 Rn01447976_m1 – Thermo Fisher Scientific) for genes of interest on a CFX Connect real-time PCR instrument (Bio-Rad).

Techniques: Knock-Out, Staining, Fluorescence, In Vivo

Journal: bioRxiv

Article Title: Pannexin 1 Channels Control Cardiomyocyte Metabolism and Neutrophil Recruitment During Non-Ischemic Heart Failure

doi: 10.1101/2023.12.29.573679

Figure Lengend Snippet: A) Quantification of uniquely expressed genes from bulk RNA-Sequencing of isolated cardiomyocytes from untreated 12-week-old male Panx1 fl/fl and Panx1 MyHC6 mice. B) Volcano plot of differentially expressed genes with a cut off of p < 0.05. Individual genes are indicated with gene name and arrow. C) Heatmap showing relative gene expression of glycolysis associated genes from isolated cardiomyocytes of untreated Panx1 fl/fl and Panx1 MyHC6 mice plotted as the Log2 (fold change) from Panx1 fl/fl mouse. Purple heatmap represents -log(p-value) of two-tailed t- tests comparing fold change of Panx1 fl/fl and Panx1 MyHC6 . (Panx1 fl/fl N = 3 mice, Panx1 MyHC6 N = 3 mice) D) Quantification of Basal extracellular acidification rate (ECAR) and E) Maximal ECAR of control of Panx1-targeted siRNA treated H9c2 cells after stimulation with either vehicle or ISO (20μM) for 1 hour. (N = 5) F) Glucose uptake as measured by 2NBD-glucose fluorescence intensity of control or Panx1-targeted siRNA treated after 1 hour ISO (20μM) stimulation. (N = 4) Data represented as mean ± SEM. Significance determined using One-way ANOVA (D, E, F) with Sidak’s multiple comparison test for post-hoc analysis for comparisons between individual groups. ns = not significant. See also S3 in reference to .

Article Snippet: Quantitative reverse transcription PCR was performed using TaqManFast Advanced Master Mix (Thermo Fisher Scientific) and TaqMan primers (mouse Panx1 mm0045091 – Invitrogen) (Rat Panx1 Rn01447976_m1 – Thermo Fisher Scientific) for genes of interest on a CFX Connect real-time PCR instrument (Bio-Rad).

Techniques: RNA Sequencing, Isolation, Gene Expression, Two Tailed Test, Control, Fluorescence, Comparison

Journal: bioRxiv

Article Title: Pannexin 1 Channels Control Cardiomyocyte Metabolism and Neutrophil Recruitment During Non-Ischemic Heart Failure

doi: 10.1101/2023.12.29.573679

Figure Lengend Snippet: A) Schematic of experimental model of isoproterenol induced non-ischemic heart failure. B) Normalized heart mass from 10–12-week-old Panx1 fl/fl or Panx1 MyHC6 mice treated for 14 days with isoproterenol (15 mg/kg/day), or saline control normalized to tibial length. (Saline: Panx1 fl/fl N = 9, Panx1 MyHC6 N = 8; Isoproterenol Panx1 fl/fl N = 11, Panx1 MyHC6 N = 9) C) Representative confocal micrographs of wheat germ agglutinin (WGA) staining at 20X (Left) and 60X (Right) from Panx1 fl/fl or Panx1 MyHC6 mice treated with isoproterenol for 14 days. Cardiomyocyte cross sectional area was calculated from WGA. (Panx1 fl/fl N = 4 mice, Panx1 MyHC6 N = 4, data points represent average of 50 cells per mouse). D) Representative M-mode images of Panx1 fl/fl mice or Panx1 MyHC6 after 14 days of isoproterenol treatment, visualized by 2D echocardiography. E) Left, paired analysis of left ventricle end-systolic volume (μL) pre- and post- isoproterenol treatment; right, percent change from baseline after isoproterenol treatment. (Panx1 fl/fl N = 8, Panx1 MyHC6 N = 9) F) Left, paired analysis of left ventricle end-diastolic volume (μL) pre- and post- isoproterenol treatment; right, percent change from baseline after isoproterenol treatment. (Panx1 fl/fl N = 8, Panx1 MyHC6 N = 9) Data represented as mean ± SEM. Significance determined using One-way ANOVA (B with Sidak’s multiple comparison test for post-hoc analysis for comparisons between individual groups; Student’s t-test (C) and Welch’s t-test (E, F) was used for comparison between two groups. Significance of paired data was determined using Mixed effects analysis (E, F) with Sidak’s multiple comparison test for post-hoc analysis for comparisons between timepoints. ns = not significant. See also S4, S5 in reference to .

Article Snippet: Quantitative reverse transcription PCR was performed using TaqManFast Advanced Master Mix (Thermo Fisher Scientific) and TaqMan primers (mouse Panx1 mm0045091 – Invitrogen) (Rat Panx1 Rn01447976_m1 – Thermo Fisher Scientific) for genes of interest on a CFX Connect real-time PCR instrument (Bio-Rad).

Techniques: Saline, Control, Staining, Comparison

Journal: bioRxiv

Article Title: Pannexin 1 Channels Control Cardiomyocyte Metabolism and Neutrophil Recruitment During Non-Ischemic Heart Failure

doi: 10.1101/2023.12.29.573679

Figure Lengend Snippet: A) Quantification of differentially expressed genes identified in bulk RNA-Sequencing between whole hearts of Panx1 fl/fl and Panx1 MyHC6 mice after 14 days of isoproterenol treatment. Differentially expressed gene determined by cut off of p < 0.05. Significantly changed, as determined by cut off of p < 0.05, KEGG Pathways and Gene Ontology (GO) Terms identified from differentially expressed genes plotted as ordered by -log(p-value). B) Heatmap demonstrating genes identified in WikiPathways “Inflammatory response”. C) Heatmap demonstrating genes identified in WikiPathways “Fatty acid oxidation and Fatty acid beta-oxidation”. D) Heatmap demonstrating genes identified in WikiPathways “Purinergic signaling”. Gradient plotted as Log2(Fold Change) of Panx1 MyHC6 compared to Panx1 fl/fl . (Panx1 fl/fl N = 5, Panx1 MyHC6 N = 4)

Article Snippet: Quantitative reverse transcription PCR was performed using TaqManFast Advanced Master Mix (Thermo Fisher Scientific) and TaqMan primers (mouse Panx1 mm0045091 – Invitrogen) (Rat Panx1 Rn01447976_m1 – Thermo Fisher Scientific) for genes of interest on a CFX Connect real-time PCR instrument (Bio-Rad).

Techniques: RNA Sequencing

Journal: bioRxiv

Article Title: Pannexin 1 Channels Control Cardiomyocyte Metabolism and Neutrophil Recruitment During Non-Ischemic Heart Failure

doi: 10.1101/2023.12.29.573679

Figure Lengend Snippet: A) Heatmap demonstrating genes identified in WikiPathway “Inflammatory response”. Gradient plotted as Log2(Fold Change) of Panx1 MyHC6 isolated cardiomyocytes after 14- days of isoproterenol treatment compared to Panx1 fl/fl . Significantly decreased, as determined by cut off of p < 0.05, GO terms in Panx1 MyHC6 cardiomyocytes compared with Panx1 fl/fl cardiomyocytes. (Panx1 fl/fl N = 4, Panx1 MyHC6 N = 4 mice). B) Flow cytometric analysis of CD45 + cells as a percentage of total live cells isolated from Panx1 fl/fl and Panx1 MyHC6 hearts after 14-days of isoproterenol treatment as identified. Representative histograms of the CD45 populations from live parent gate. (Panx1 fl/fl N = 8, Panx1 MyHC6 N = 8 mice) C) Flow cytometric analysis of neutrophils (CD11b + , Ly6g + ) as a percentage of the CD45 + cells in Panx1 fl/fl and Panx1 MyHC6 hearts after 14-days of isoproterenol treatment. Representative plots of the identified neutrophils from Panx1 fl/fl and Panx1 MyHC6 mice hearts. (Panx1 fl/fl N = 8, Panx1 MyHC6 N = 8 mice) D) Representative confocal micrographs of Ly6g staining at 60X from Panx1 fl/fl and Panx1 MyHC6 mice treated with isoproterenol for 14 days. Ly6g was used to stain neutrophils; DAPI was used as a nuclear stain. Quantification of number of neutrophils, Ly6g + puncta, per 60X field in whole heart tissue from Panx1 fl/fl and Panx1 MyHC6 mice after 14-days of isoproterenol treatment. (Panx1 fl/fl N = 8, Panx1 MyHC6 N = 9 mice, 3-5 images per mouse averaged) E) Flow cytometric analysis of macrophages (CD11b + , F4/80 + ) as a percentage of CD45 + cells from Panx1 fl/fl and Panx1 MyHC6 hearts after 14-days of isoproterenol treatment. Macrophages expressing the co-stimulatory activation marker (CD86 + ) and macrophages expressing the anti-inflammatory marker (CD163 + , CD86 - ) as a percentage of the macrophages (CD11b + , F4/80 + ). Flow cytometric analysis of dendritic cells (DC) (CD11b + , CD24 + ) as a percentage of CD45 + cells from Panx1 fl/fl and Panx1 MyHC6 hearts after 14-days of isoproterenol treatment. (Panx1 fl/fl N = 6, Panx1 MyHC6 N = 6 mice) Data represented as mean ± SEM. Significance determined using Welch’s t-test (B, C, D, E) was used for comparison between two groups. ns = not significant. See also S6 in reference to .

Article Snippet: Quantitative reverse transcription PCR was performed using TaqManFast Advanced Master Mix (Thermo Fisher Scientific) and TaqMan primers (mouse Panx1 mm0045091 – Invitrogen) (Rat Panx1 Rn01447976_m1 – Thermo Fisher Scientific) for genes of interest on a CFX Connect real-time PCR instrument (Bio-Rad).

Techniques: Isolation, Staining, Expressing, Activation Assay, Marker, Comparison