anti α sma  (Proteintech)

Journal: Pain

Article Title: The prokineticin system and glia cells as pharmacological targets to control neuroinflammation and to relieve pain in a murine model of Fabry–Anderson disease

doi: 10.1097/j.pain.0000000000003818

Figure Lengend Snippet: Sciatic nerve: mRNA levels of PKS and neuroinflammatory markers and effects of PC1 and minocycline treatments. mRNA expression levels (RT-qPCR) of the PKS members (A) PK2, (B) PKR1 and (C) PKR2, proinflammatory cytokines (D) IL-6, (E) IL-1β, and (F) TNF-α, (G) macrophage marker Iba1 and (H) Schwann cell marker GFAP were evaluated in the sciatic nerve (after 14 days of pharmacological treatment). Results are normalized to the housekeeping gene GAPDH and expressed as fold over the age-matched CTR group. Data are the mean ± SD of 4 to 6 animals/group. Statistical analyses were performed by one-way ANOVA followed by the Šidák post hoc test. * P < 0.05, ** P < 0.01, *** P < 0.001 vs age-matched CTR; ◦ P < 0.05, ○○ P < 0.01, ○○○ P < 0.001 vs age-matched FD; ++ P < 0.01 vs age-matched FD + PC1. Treatments: (A) 10 weeks: F (3,20) = 7.912, P = 0.0011; 25 weeks: F (3,12) = 1.745, P = 0.2110 (B) 10 weeks: F (3,20) = 18.91, P < 0.0001; 25 weeks: F (3,20) = 1.077, P = 0.3814 (C) 10 weeks: F (3,20) = 1.559, P = 0.2305; 25 weeks: F (3,12) = 1.005, P = 0.4243 (D) 10 weeks: F (3,20) = 7.587, P = 0.0014; 25 weeks: F (3,20) = 0.1821, P = 0.9073 (E) 10 weeks: F (3,20) = 6.204, P = 0.0037; 25 weeks: F (3,20) = 0.1870, P = 0.9040 (F) 10 weeks: F (3,20) = 4.867, P = 0.0106; 25 weeks: F (3,20) = 6.058, P = 0.0042 (G) 10 weeks: F (3,20) = 4.306, P = 0.0169; 25 weeks: F (3,20) = 4.075, P = 0.0207 (H) 10 weeks: F (3,20) = 27.75, P < 0.0001; 25 weeks: F (3,20) = 1.119, P = 0.3648. ANOVA, analysis of variance; FD, Fabry–Anderson disease; IL, interleukin; PKR, prokineticin receptors; PKS, Prokineticin System; TNF-α, tumor necrosis factor-α.

Article Snippet: Real-time PCR was executed using Luna Universal Probe, qPCR Master Mix (BioLabs), and specific Taqman Gene expression assays (Thermofisher Scientific, Waltham, MA) as prokineticin 2 (PK2: Mm01182450_g1), prokineticin receptors (PKR1: Mm00517546_m1 and PKR2: Mm00769571_m1), interleukin 6 (IL-6: Mm00446190_m1), interleukin-1β (IL-1β: Mm00434228_m1), tumor necrosis factor-α (TNF-α: Mm00443258_m1), glial fibrillary acidic protein (GFAP: Mm01253033_m1), ionized calcium-binding adapter molecule 1 (Iba1: Mm00479862_g1), and Glyceraldehyde-3-phosphate dehydrogenase (GAPDH:Mm99999915_g1).

Techniques: Expressing, Quantitative RT-PCR, Marker

Journal: Pain

Article Title: The prokineticin system and glia cells as pharmacological targets to control neuroinflammation and to relieve pain in a murine model of Fabry–Anderson disease

doi: 10.1097/j.pain.0000000000003818

Figure Lengend Snippet: Dorsal root ganglia: mRNA levels of PKS, neuroinflammatory, epigenetic regulators and effects of PC1 and minocycline treatments. mRNA expression levels (RT-qPCR) of the PKS members (A) PK2, (B) PKR1 and (C) PKR2, proinflammatory cytokines (D) IL-6, (E) IL-1β, and (F) TNF-α, (G) macrophages marker Iba1, (H) satellite glial cell marker GFAP, (I) PPARγ, (J) KDM6A and (K) KDM6B were evaluated in the dorsal root ganglia (after 14 days of pharmacological treatment). Results are normalized to the housekeeping gene GAPDH and expressed as fold over the age-matched CTR group. Data are (A–H) the mean ± SD of 4 to 6 animals/group. Statistical analyses were performed by 1-way ANOVA followed by the Šidák post hoc test. * P < 0.05, ** P < 0.01, *** P < 0.001 vs age-matched CTR; ◦ P < 0.05, ○○ P < 0.01, ○○○ P < 0.001 vs age-matched FD; + P < 0.05 vs age-matched FD + PC1. Treatments: (A) 10 weeks: F (3,20) = 8.931, P = 0.0006; 25 weeks: F (3,20) = 10.22, P = 0.0003 (B) 10 weeks: F (3,20) = 0.5349, P = 0.6636; 25 weeks: F (3,20) = 0.5671, P = 0.6430 (C) 10 weeks: F (3,20) = 2.232, P = 0.1158; 25 weeks: F (3,20) = 1.015, P = 0.4069 (D) 10 weeks: F (3,20) = 2.239, P = 0.1150; 25 weeks: F (3,20) = 6.236, P = 0.0037 (E) 10 weeks: F (3,20) = 5.132, P = 0.0086; 25 weeks: F (3,20) = 4.947, P = 0.0099 (F) 10 weeks: F (3,20) = 7.602, P = 0.0014; 25 weeks: F (3,20) = 7.381, P = 0.0016 (G) 10 weeks: F (3,20) = 0.5217, P = 0.6723; 25 weeks: F (3,20) = 7.345, P = 0.0017 (H) 10 weeks: F (3,20) = 11.61, P = 0.0001; 25 weeks: F (3,20) = 1.768, P = 0.1856 (I) 10 weeks: F (3,14) = 5.393, P = 0.0112; 25 weeks: F (3,19) = 4.826, P = 0.0116 (J) 10 weeks: F (3,16) = 7.005, P = 0.0032; 25 weeks: F (3,12) = 0.9353, P = 0.4538 (K) 10 weeks: F (3,16) = 0.4278, P = 0.7358; 25 weeks: F (3,19) = 3.613, P = 0.322. FD, Fabry–Anderson disease; IL, interleukin; PKR, prokineticin receptors; PKS, Prokineticin System; TNF-α, tumor necrosis factor-α.

Article Snippet: Real-time PCR was executed using Luna Universal Probe, qPCR Master Mix (BioLabs), and specific Taqman Gene expression assays (Thermofisher Scientific, Waltham, MA) as prokineticin 2 (PK2: Mm01182450_g1), prokineticin receptors (PKR1: Mm00517546_m1 and PKR2: Mm00769571_m1), interleukin 6 (IL-6: Mm00446190_m1), interleukin-1β (IL-1β: Mm00434228_m1), tumor necrosis factor-α (TNF-α: Mm00443258_m1), glial fibrillary acidic protein (GFAP: Mm01253033_m1), ionized calcium-binding adapter molecule 1 (Iba1: Mm00479862_g1), and Glyceraldehyde-3-phosphate dehydrogenase (GAPDH:Mm99999915_g1).

Techniques: Expressing, Quantitative RT-PCR, Marker

Journal: Pain

Article Title: The prokineticin system and glia cells as pharmacological targets to control neuroinflammation and to relieve pain in a murine model of Fabry–Anderson disease

doi: 10.1097/j.pain.0000000000003818

Figure Lengend Snippet: Spinal cord: mRNA levels of PKS and neuroinflammatory markers and effects of PC1 and minocycline. mRNA expression levels (RT-qPCR) of the PKS members (A) PK2, (B) PKR1 and (C) PKR2, proinflammatory cytokines (D) IL-6, (E) IL-1β, and (F) TNF-α, (G) microglia marker Iba1 and (H) astrocytes marker GFAP were evaluated in the spinal cord (after 14 days of pharmacological treatment). Results are normalized to the housekeeping gene GAPDH and expressed as fold over the age-matched CTR group. Data are the mean ± SD of 6 animals/group. Statistical analyses were performed by 1-way ANOVA followed by the Šidák post hoc test. * P < 0.05 vs age-matched CTR; ○○ P < 0.01 vs age-matched FD. Treatments: (A) 10 weeks: F (3,20) = 0.3411, P = 0.9913; 25 weeks: F (3,20) = 0.5229, P = 0.6715 (B) 10 weeks: F (3,20) = 0.9708, P = 0.4260; 25 weeks: F (3,20) = 0.3878, P = 0.7630 (C) 10 weeks: F (3,20) = 0.4009, P = 0.7539; 25 weeks: F (3,20) = 1.428, P = 0.2642 (D) 10 weeks: F (3,20) = 2.787, P = 0.0672; 25 weeks: F (3,20) = 7.518, P = 0.0015 (E) 10 weeks: F (3,20) = 3.406, P = 0.0376; 25 weeks: F (3,20) = 0.6931, P = 0.5670 (F) 10 weeks: F (3,20) = 3.981, P = 0.0224; 25 weeks: F (3,20) = 0.6343, P = 0.6016 (G) 10 weeks: F (3,20) = 1.136, P = 0.2824; 25 weeks: F (3,20) = 8.352, P = 0.0008 (H) 10 weeks: F (3,20) = 3.490, P = 0.0348; 25 weeks: F (3,20) = 6.563, P = 0.0029. ANOVA, analysis of variance; Iba1, ionized calcium-binding adapter molecule 1; IL, interleukin; PKR, prokineticin receptors; PKS, Prokineticin System; TNF-α, tumor necrosis factor-α.

Article Snippet: Real-time PCR was executed using Luna Universal Probe, qPCR Master Mix (BioLabs), and specific Taqman Gene expression assays (Thermofisher Scientific, Waltham, MA) as prokineticin 2 (PK2: Mm01182450_g1), prokineticin receptors (PKR1: Mm00517546_m1 and PKR2: Mm00769571_m1), interleukin 6 (IL-6: Mm00446190_m1), interleukin-1β (IL-1β: Mm00434228_m1), tumor necrosis factor-α (TNF-α: Mm00443258_m1), glial fibrillary acidic protein (GFAP: Mm01253033_m1), ionized calcium-binding adapter molecule 1 (Iba1: Mm00479862_g1), and Glyceraldehyde-3-phosphate dehydrogenase (GAPDH:Mm99999915_g1).

Techniques: Expressing, Quantitative RT-PCR, Marker, Binding Assay

Journal: Pain

Article Title: The prokineticin system and glia cells as pharmacological targets to control neuroinflammation and to relieve pain in a murine model of Fabry–Anderson disease

doi: 10.1097/j.pain.0000000000003818

Figure Lengend Snippet: Spinal cord: GFAP protein levels, effects of PC1 and minocycline and qualitative immunohistochemistry. Protein expression levels (Western Blot) of the astrocyte marker GFAP (A) in young (10-week-old) and (B) adult (25-week-old) mice were evaluated in the spinal cord (after 14 days of pharmacological treatment). Results are normalized to the housekeeping protein β-actin and expressed as fold over the age-matched CTR group. Data are the mean ± SD of 6 animals/group. Statistical analyses were performed by 1-way ANOVA followed by the Šidák post hoc test. * P < 0.05 vs age-matched CTR. Treatments: (A) 10 weeks: F (3,20) = 3.956, P = 0.0229 (B) 25 weeks: F (3,20) = 3.955, P = 0.0230. (C) Immunohistochemistry qualitative images of GFAP signal in the spinal cord are provided. Scale bars: 50 μm. ANOVA, analysis of variance.

Article Snippet: Real-time PCR was executed using Luna Universal Probe, qPCR Master Mix (BioLabs), and specific Taqman Gene expression assays (Thermofisher Scientific, Waltham, MA) as prokineticin 2 (PK2: Mm01182450_g1), prokineticin receptors (PKR1: Mm00517546_m1 and PKR2: Mm00769571_m1), interleukin 6 (IL-6: Mm00446190_m1), interleukin-1β (IL-1β: Mm00434228_m1), tumor necrosis factor-α (TNF-α: Mm00443258_m1), glial fibrillary acidic protein (GFAP: Mm01253033_m1), ionized calcium-binding adapter molecule 1 (Iba1: Mm00479862_g1), and Glyceraldehyde-3-phosphate dehydrogenase (GAPDH:Mm99999915_g1).

Techniques: Immunohistochemistry, Expressing, Western Blot, Marker