Journal: Developmental Dynamics

Article Title: The endocannabinoid system regulates both ependymoglial and neuronal cell responses to a tail amputation in the axolotl

doi: 10.1002/dvdy.70035

Figure Lengend Snippet: Identification and conservation of the axolotl endocannabinoid receptors. (A) Protein sequence alignment of the putative axolotl CB1 sequence with the rat and zebrafish CB1 sequence. (B) Protein sequence alignment of the putative axolotl CB2 sequence with the rat and zebrafish CB2 sequence. Red asterisks and red boxes indicate amino acids that are conserved between all three species. (C) Western blot using the rat CB1 antibody on axolotl tail tissue demonstrates a single prominent band at ~120 kDa ( n = 3). (D) Western blot using the rat CB2 antibody on axolotl tail tissue demonstrates two bands at a similar molecular weight of ~46 kDa ( n = 3). MW = molecular weight (for each band in ladder). (E) Preadsorption control for the CB1 antibody using either CB1 or CB2 antigenic peptides ( n = 3). (F) Preadsorption control for the CB2 antibody using CB1 or CB2 antigenic peptides ( n = 3).

Article Snippet: Primary antibodies included CB1 (1:100, Alomone Labs), CB2 (1:100, Alomone Labs), GFAP (1:100, Chemicon), NeuN (1:100, Chemicon), β‐III‐tubulin (1:500, Sigma), and DCX (1:50, DSHB).

Techniques: Sequencing, Western Blot, Molecular Weight, Control

Journal: Developmental Dynamics

Article Title: The endocannabinoid system regulates both ependymoglial and neuronal cell responses to a tail amputation in the axolotl

doi: 10.1002/dvdy.70035

Figure Lengend Snippet: CB1 and CB2 are upregulated in response to tail amputation. (A) Western blot analysis demonstrates a significant upregulation of CB1 in the first 3 days after tail amputation, compared to uninjured controls ( n = 3; F (4,40) = 5.994, p = .0007, one‐way ANOVA). (B) No change in CB2 expression is shown in the first 3 days post tail amputation ( n = 3; F (4,40) = 2.779, p = .0397, one‐way ANOVA). (C) Western blot analysis demonstrates a significant upregulation of CB1 expression at both 7 and 14 days after tail amputation ( n = 3; F (2,24) = 15.97, p < .0001, one‐way ANOVA). (D) Western blot analysis demonstrates a significant upregulation of CB2 at 7 and 14 days post tail amputation ( n = 3; F (2,24) = 10.84, p = .0004, one‐way ANOVA). Uninj = uninjured tail tissue. hpa = hours post tail amputation; dpa = days post tail amputation. ns = not significant. * p < .05, ** p < .01, *** p < .001, *** *p < .0001 compared to uninjured controls. # p < .05.

Article Snippet: Primary antibodies included CB1 (1:100, Alomone Labs), CB2 (1:100, Alomone Labs), GFAP (1:100, Chemicon), NeuN (1:100, Chemicon), β‐III‐tubulin (1:500, Sigma), and DCX (1:50, DSHB).

Techniques: Western Blot, Expressing

Journal: Developmental Dynamics

Article Title: The endocannabinoid system regulates both ependymoglial and neuronal cell responses to a tail amputation in the axolotl

doi: 10.1002/dvdy.70035

Figure Lengend Snippet: CB1 and CB2 are expressed in ependymoglia and neurons in the regenerating spinal cord. (A) Schematic displays the cell‐type architecture of the axolotl spinal cord. The spinal cord is comprised of ependymoglial cells (blue) that line the central canal (cc) of the spinal cord. These ependymoglia extend GFAP + processes toward the periphery of the spinal cord. The spinal cord also contains NeuN + neurons (green) that surround the ependymoglia and extend axons that express β‐III‐tubulin. (B) Immunohistochemistry ( n = 3) shows the absence of CB1 from neuronal cell bodies (iv), and shows the co‐localization of CB1 with β‐III‐tubulin in axons (viii, yellow arrow) and with GFAP in glial cell processes (xii, blue arrow). (C) Immunohistochemistry ( n = 3) shows the absence of CB2 from neuronal cell bodies (iv), and displays the co‐localization of CB2 with β‐III‐tubulin in axons (viii, yellow arrow) and with GFAP in glial cells (xii, blue arrow). (D) Fluorescent in situ hybridization ( n = 2) demonstrates cb1 mRNA expression in both neurons (yellow arrow) and ependymoglia (blue arrow). Scale bars: 100 μm.

Article Snippet: Primary antibodies included CB1 (1:100, Alomone Labs), CB2 (1:100, Alomone Labs), GFAP (1:100, Chemicon), NeuN (1:100, Chemicon), β‐III‐tubulin (1:500, Sigma), and DCX (1:50, DSHB).

Techniques: Immunohistochemistry, In Situ Hybridization, Expressing

Journal: Developmental Dynamics

Article Title: The endocannabinoid system regulates both ependymoglial and neuronal cell responses to a tail amputation in the axolotl

doi: 10.1002/dvdy.70035

Figure Lengend Snippet: Inhibiting CB1 and CB2 receptor signaling impairs tail regeneration. (A) Representative images of tail regenerates after a 7‐day treatment with the vehicle (control, i), 1 μM AM251 (ii), or 1 μM AM630 (iii). Black dotted line indicates the original plane of amputation. Scale bar: 1 mm. (B, C) Graphs show that the proportional increase in axolotl body length was significantly reduced following either a 7‐day treatment with either 1 μM AM251 ( n = 8; B) or after a 7‐day treatment with 1 μM AM630 ( n = 8; C) compared to the vehicle control (unpaired t tests). (D) Graph shows a significant reduction in the proportional increase in axolotl body length (7 days after tail amputation) following only a 1‐day pulse treatment with either 1 μM AM251 ( n = 10) or 1 μM AM630 ( n = 10), compared to vehicle controls ( n = 10; F (2,27) = 18.86; p < .0001, one‐way ANOVA). (E) Western blot analyses show that treatment with AM251 prevented the upregulation of CB1 that normally occurs in untreated or vehicle‐treated control animals at 7‐days post tail amputation ( n = 3; Constant 7‐day bath treatment: F (3,32) = 14.69; p < .0001; 1‐day pulse treatment: F (3,32) = 18.60; p < .0001; one‐way ANOVAs). Representative blot for 1‐day pulse treatment shown. (F) Treatment with AM630 prevented the upregulation of CB2 that normally occurs in untreated or vehicle‐treated control animals at 7‐days post tail amputation ( n = 3; constant treatment: F (3,32) = 24.80; p < .0001; 1‐day pulse treatment: F (3,32) = 11.60; p < .0001; one‐way ANOVAs). Representative blot for 7‐day constant treatment shown. * *p < .01, ** *p < .001, *** *p < .0001 compared to vehicle controls. ### p < .001. #### p < .0001.

Article Snippet: Primary antibodies included CB1 (1:100, Alomone Labs), CB2 (1:100, Alomone Labs), GFAP (1:100, Chemicon), NeuN (1:100, Chemicon), β‐III‐tubulin (1:500, Sigma), and DCX (1:50, DSHB).

Techniques: Control, Western Blot

Journal: Developmental Dynamics

Article Title: The endocannabinoid system regulates both ependymoglial and neuronal cell responses to a tail amputation in the axolotl

doi: 10.1002/dvdy.70035

Figure Lengend Snippet: Inhibiting cannabinoid receptor activity reduces ependymoglial cell proliferation and upregulates GFAP + in glial cell processes. (A) Representative images of EdU + cells in the regenerating axolotl spinal cord at 7‐days post tail amputation after treatment with 1 μM AM251 (ii), 1 μM AM630 (iii), or the vehicle (control, i). White dotted circles outline the spinal cord. (B) Graph shows a significant reduction in the proportion of EdU + cells in the axolotl spinal cord at 7‐days post tail amputation after treatment with either 1 μM AM251 ( n = 4) or 1 μM AM630 ( n = 4) in comparison to vehicle controls ( n = 4; F (2,9) = 25.25; p = .0002, one‐way ANOVA). ** *p < .001 compared to vehicle controls. (C) Representative images of GFAP expression in uninjured axolotl tail tissue (i) and in regenerating tail tissue (ii) at 7‐days post tail amputation (dpa). (D) Quantified western blot data demonstrates a significant reduction in GFAP expression in the first 7‐days post tail amputation in comparison to uninjured tail tissue ( n = 3; F (3,32) = 25.97, p < .0001, one‐way ANOVA). ** *p < .001 compared to uninjured controls. (E, F) Immunohistochemistry shows GFAP expression paired with either CB1 (E) or CB2 (F) staining in the axolotl spinal cord at 7‐days post tail amputation after treatment with 1 μM AM251 (Eii), or 1 μM AM630 (Fii) or the vehicle (controls, Ei and Fi). Scale bars = 100 μm.

Article Snippet: Primary antibodies included CB1 (1:100, Alomone Labs), CB2 (1:100, Alomone Labs), GFAP (1:100, Chemicon), NeuN (1:100, Chemicon), β‐III‐tubulin (1:500, Sigma), and DCX (1:50, DSHB).

Techniques: Activity Assay, Control, Comparison, Expressing, Western Blot, Immunohistochemistry, Staining

Journal: Cells

Article Title: Muramyl Dipeptide Administration Delays Alzheimer’s Disease Physiopathology via NOD2 Receptors

doi: 10.3390/cells11142241

Figure Lengend Snippet: List of antibodies used for the Western blot and immunostaining analyses and all related information, including the name of the company, molecular weight, species, primary and secondary antibody dilution, and targets.

Article Snippet: ABCB1 , Novus biological , 170 kDa , Rabbit , NBP2-67667 , 1/1000 , IgG anti-Rabbit peroxidase , 111-035-144 , 1/2000.

Techniques: Western Blot, Immunostaining, Molecular Weight, Concentration Assay

Journal: Cells

Article Title: Muramyl Dipeptide Administration Delays Alzheimer’s Disease Physiopathology via NOD2 Receptors

doi: 10.3390/cells11142241

Figure Lengend Snippet: The long-term administration of MDP has a different effect on amyloid regarding the sex. Although, MDP induces an increase in the plaque number in the female APP Swe /PS1 treatment with MDP group. ( A ): Unbiased stereological analysis of the 6E10 + plaque number of the male and female APP Swe /PS1 treatment or not with MDP groups in the hippocampus and cortex. Analysis showed a diminution in the plaque number in the male APP Swe /PS1 treatment group in both areas and an augmentation in the Aβ plaque numbers in the female APP Swe /PS1 treatment group in the hippocampus compared to sex-matched APP Swe /PS1 controls. ( B ): Average volume per 6E10 + plaque in µm 3 of the male and female APP Swe /PS1 treatment or not groups with MDP in the hippocampus and cortex. Analysis showed no difference in both sexes and area compared to sex-matched APP Swe /PS1 controls. ( C ): The 6E10+ blood vessel frequency of male and female APP Swe /PS1 treatment or not with MDP groups. Analysis showed an augmentation in the male APP Swe /PS1 treatment group but no difference in the female APP Swe /PS1 treatment groups compared to sex-matched APP Swe /PS1 controls. ( D ): Elisa Aβ40 and Aβ42 of the male and female APP Swe /PS1 treatment or not with MDP groups. Analysis showed an increase in Aβ40 in the male APP Swe /PS1 treatment group but a decrease in the Aβ42 in APP Swe /PS1 treatment group for both sexes compared to sex-matched APP Swe /PS1 controls. ( E ): Ratio of Aβ42/Aβ40 in the male and female APP Swe /PS1 mice treatment or not with MDP groups. Analysis showed a diminution in the male APP Swe /PS1 treatment group but no difference in the female APP Swe /PS1 treatment group compared to sex-matched APP Swe /PS1 controls. ( F ); ( G ): MDP increases the clearance systems through LRP1 and ABCB1 expression in the male group. Western blot analysis of ( F ) LRP1 ( G ) ABCB1 in the male and female wild-type and APP Swe /PS1 treatment or not with MDP groups. Analysis showed an augmentation in LRP1 and ABCB1 expression in the male APP Swe /PS1 treatment group but a diminution in LRP1 and ABCB1 expression in the female APP Swe /PS1 treatment group compared to sex-matched APP Swe /PS1 controls. Data are mean ± SEM ( n = 8 animals/group) * p < 0.05, *** p < 0.001 compared to sex-matched APP Swe /PS1 controls.

Article Snippet: ABCB1 , Novus biological , 170 kDa , Rabbit , NBP2-67667 , 1/1000 , IgG anti-Rabbit peroxidase , 111-035-144 , 1/2000.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Western Blot

Journal: Frontiers in bioscience (Landmark edition)

Article Title: PCAT1/miR-129/ABCB1 axis confers chemoresistance in non-small cell lung cancer.

doi: 10.2741/4842

Figure Lengend Snippet: Figure 4. ABCB1 expression upregulated by PCAT1/miR-129 axis contributes to DDP resistance in A549/DDP and H1299/DDP cells. (A) The predicted binding site of miR-129 within the ABCB1 3’ UTR and the mutated sites are shown. (B) Luciferase reporter assay of 293T cells co- transfected with reporter constructs (ABCB1-WT or ABCB1-MUT) and miR-con, miR-129, vector, or pcDNA-PCAT1. (C) qRT-PCR assay was performed to analyze the expression of miR-129 in A549/DDP and H1299/DDP cells transfected with miR-con, miR-129, si-con, si-PCAT1#2, or anti-miR-129. (D and E) Western blot analysis of miR-129 expression in A549/DDP and H1299/DDP cells transfected with miR-con, miR- 129, si-con, si-PCAT1#2, or anti-miR-129. (F) qRT-PCR analysis showed that transfection with si-PCAT1#2 downregulated the expression of ABCB1 in A549/DDP and H1299/DDP cells. (G) IC50 values of DDP in si-con- and si-ABCB1-transfected cells. (H) Flow cytometry analysis of apoptosis in si-con- and si-ABCB1-transfected cells. *P < 0.05.

Article Snippet: Subsequently, membranes were rinsed with tris-buffered saline with tween, blocked for 1 h in 951 © 1996-2020 non-fat milk, and then probed with special primary antibody against ABCB1 (Novus Biologicals, Littleton, CO, USA) overnight at 4°C.

Techniques: Expressing, Binding Assay, Luciferase, Reporter Assay, Transfection, Construct, Plasmid Preparation, Quantitative RT-PCR, Western Blot, Flow Cytometry