Journal: Cell Death and Differentiation

Article Title: APP upregulation contributes to retinal ganglion cell degeneration via JNK3

doi: 10.1038/s41418-017-0005-3

Figure Lengend Snippet: a Pathway analysis of microarray data on genes differentially expressed between control and SH-SY5Y-APP cells. b ChIP-seq screening results in SH-SY5Y cells showed that the region of AICD interaction is in the fourth intron of the JNK3 locus in chromosome 4. TSS, transcription starting site. c Validation of the ChIP-seq results. SH-SY5Y cells were transfected with either pcDNA4/V5-His or pcDNA4/V5-His-hAICD59 plasmid, and subjected to ChIP assay, using normal mouse IgG as control. Input, 5% of the sonicated chromatin. Left panel, real-time PCR results show V5, but not IgG control interacts with the JNK3 intron region ( n = 3). Right panel, representative image shows the real-time PCR products resolved by agarose gel. The molecular weight of the DNA ladder shown in bp on the left. d Real-time PCR results showing the JNK3 mRNA level in pcDNA4/V5-His (control) or pcDNA4/V5-His-hAICD59 (AICD) transfected SH-SY5Y cells ( n = 3). Student’s t- test: ** p < 0.01. Error bars represent SEM. e Luciferase assay showing the enhancer activity of the AICD-interacting site. One-way ANOVA followed by post-hoc Bonferroni test: ** p < 0.01. Error bars represent SEM

Article Snippet: Rabbit antibodies against APP (Y188, Abcam) [ ], JNK1 (catalog no. MAB17761, R&D Systems), JNK2 (catalog no. MAB1846, R&D Systems) [ ], JNK3 (catalog no. 2305, Cell Signaling Technology), p-cJun73 (catalog no. 9164, Cell Signaling Technology) and c-Jun (catalog no. 9165, Cell Signaling Technology); mouse antibodies against GAPDH (catalog no. ab8245, Abcam) [ ], V5 (catalog no. R96025, Invitrogen), pJNK (catalog no. 9255, Cell Signaling Technology) [ ] and JNK3 (catalog no. CP10162, Cell Applications) were used.

Techniques: Microarray, Control, ChIP-sequencing, Biomarker Discovery, Transfection, Plasmid Preparation, Sonication, Real-time Polymerase Chain Reaction, Agarose Gel Electrophoresis, Molecular Weight, Luciferase, Activity Assay

Journal: Cell Death and Differentiation

Article Title: APP upregulation contributes to retinal ganglion cell degeneration via JNK3

doi: 10.1038/s41418-017-0005-3

Figure Lengend Snippet: a Intact retinae from wild-type mice were fixed, paraffin-embedded and 5 μm thick longitudinal sections were cut. The sections were stained with anti-JNK3, anti-APP and anti-Tuj1 antibodies after antigen retrieval. The inserts show lower magnification photomicrographs illustrating the immunofluorescence signal distribution across all the cell layers of the retina. APP and JNK3 were co-expressed in Tuj1 + RGCs. b Intact retinae from wild-type, APP-null or JNK3-null mice were stained with anti-APP or anti-JNK3 antibodies after antigen retrieval. The inserts show lower magnification photomicrographs illustrating the immunofluorescence signal distribution across all the cell layers of the retina. The anti-APP and anti-JNK3 antibodies were specific as no immunofluorescence signal is seen in the APP-null and JNK3-null retinal sections, respectively. c Adult, wild-type mouse retinae were retrogradely labeled with OHSt, fixed, and 5 μm thick sections were cut. The sections were stained with anti-APP and anti-JNK3 antibodies without antigen retrieval as antigen retrieval destroys the OHSt signal. Arrow heads indicate several OHSt-labeled RGCs, which co-express both APP and JNK3. Scale bar, 20 μm

Article Snippet: Rabbit antibodies against APP (Y188, Abcam) [ ], JNK1 (catalog no. MAB17761, R&D Systems), JNK2 (catalog no. MAB1846, R&D Systems) [ ], JNK3 (catalog no. 2305, Cell Signaling Technology), p-cJun73 (catalog no. 9164, Cell Signaling Technology) and c-Jun (catalog no. 9165, Cell Signaling Technology); mouse antibodies against GAPDH (catalog no. ab8245, Abcam) [ ], V5 (catalog no. R96025, Invitrogen), pJNK (catalog no. 9255, Cell Signaling Technology) [ ] and JNK3 (catalog no. CP10162, Cell Applications) were used.

Techniques: Staining, Immunofluorescence, Labeling

Journal: Cell Death and Differentiation

Article Title: APP upregulation contributes to retinal ganglion cell degeneration via JNK3

doi: 10.1038/s41418-017-0005-3

Figure Lengend Snippet: a Quantitative analysis of relative mRNA levels of Jnk s in intact retinae or retinae 1 day (1 d), 1 week (1 w) or 2 weeks (2 w) after ONA ( n = 3 for each time point). b Representative western blots from at least three biological repeats showing JNK expression after ONA. Intact retina or retinae 1 d, 1 w, 2 w after ONA were harvested, lysed and immunoblotted using antibodies against JNK1, JNK2, JNK3, pJNK or GAPDH. The location of molecular weight markers is shown in kDa on the right. c Representative fluorescence microscope images show immunostaining of mouse retina flat-mounts from wild-type or JNK1, JNK2, or JNK3-null mice ( n = 6 for each genotype) 14 days after ONA using an anti-Tuj1 antibody. d The bar chart shows quantitative analysis of Tuj1 + RGC numbers 14 days after ONA. Scale bar, 50 μm. One-way ANOVA followed by post-hoc Bonferroni test: ** p < 0.01. Error bars represent SEM

Article Snippet: Rabbit antibodies against APP (Y188, Abcam) [ ], JNK1 (catalog no. MAB17761, R&D Systems), JNK2 (catalog no. MAB1846, R&D Systems) [ ], JNK3 (catalog no. 2305, Cell Signaling Technology), p-cJun73 (catalog no. 9164, Cell Signaling Technology) and c-Jun (catalog no. 9165, Cell Signaling Technology); mouse antibodies against GAPDH (catalog no. ab8245, Abcam) [ ], V5 (catalog no. R96025, Invitrogen), pJNK (catalog no. 9255, Cell Signaling Technology) [ ] and JNK3 (catalog no. CP10162, Cell Applications) were used.

Techniques: Western Blot, Expressing, Molecular Weight, Fluorescence, Microscopy, Immunostaining

Journal: Cell Death and Differentiation

Article Title: APP upregulation contributes to retinal ganglion cell degeneration via JNK3

doi: 10.1038/s41418-017-0005-3

Figure Lengend Snippet: Representative western blots show JNK3 expression in a pcDNA3-FLAG-APP695 transfected or pcDNA3 transfected SH-SY5Y cells, e retinae from wild-type or APP-null mice, or i pcDNA4/V5-His-AICD59 or pcDNA4/V5-His transfected SH-SY5Y cells. The samples were lysed, resolved in SDS-page gel, and immunoblotted using antibodies against JNK3 or GAPDH. b , f , j The bar charts show quantitative analysis of relative JNK3 expression ( n = 3 for each experiment). Representative western blots show phosphorylative activation of JNK3 in c APP695-overexpressing or normal SH-SY5Y cells, g retinae from wild-type or APP-null mice, or k pcDNA4/V5-His-AICD59 or pcDNA4/V5-His transfected SH-SY5Y cells. JNK3 was immunoprecipitated with anti-JNK3 antibody and then detected for kinase activity with anti-phospho-JNK (pJNK) or anti-JNK3 antibody. Whole-cell lysate western blots show the downstream substrate pc-Jun73 or c-Jun level. d , h , l The bar charts show quantitative analysis of activated pJNK/mg lyate proteins or pc-Jun73/c-Jun ratio ( n = 3 for each experiment). Student’s t -test: * p < 0.05; ** p < 0.01. Error bars represent SEM

Article Snippet: Rabbit antibodies against APP (Y188, Abcam) [ ], JNK1 (catalog no. MAB17761, R&D Systems), JNK2 (catalog no. MAB1846, R&D Systems) [ ], JNK3 (catalog no. 2305, Cell Signaling Technology), p-cJun73 (catalog no. 9164, Cell Signaling Technology) and c-Jun (catalog no. 9165, Cell Signaling Technology); mouse antibodies against GAPDH (catalog no. ab8245, Abcam) [ ], V5 (catalog no. R96025, Invitrogen), pJNK (catalog no. 9255, Cell Signaling Technology) [ ] and JNK3 (catalog no. CP10162, Cell Applications) were used.

Techniques: Western Blot, Expressing, Transfection, SDS Page, Activation Assay, Immunoprecipitation, Activity Assay

Journal: Cell Death and Differentiation

Article Title: APP upregulation contributes to retinal ganglion cell degeneration via JNK3

doi: 10.1038/s41418-017-0005-3

Figure Lengend Snippet: a Representative western blots show JNK3 expression and overall phosphorylative activation of JNK in retinae from wild-type or APP-null mice 1 day (1 d), 1 week (1 w) and 2 weeks (2 w) after ONA ( n = 3 for each time point). Anti-JNK3, anti-pJNK and anti-GAPDH antibodies were used to detect the respective protein expression. b The bar charts show quantitative analysis of relative JNK3 expression or overall phosphorylative activation of JNK in the ipsilateral-injured retinae vs. the contralateral-uninjured retina. One-way ANOVA followed by post-hoc Bonferroni test: ** p < 0.01. Error bars represent SEM

Article Snippet: Rabbit antibodies against APP (Y188, Abcam) [ ], JNK1 (catalog no. MAB17761, R&D Systems), JNK2 (catalog no. MAB1846, R&D Systems) [ ], JNK3 (catalog no. 2305, Cell Signaling Technology), p-cJun73 (catalog no. 9164, Cell Signaling Technology) and c-Jun (catalog no. 9165, Cell Signaling Technology); mouse antibodies against GAPDH (catalog no. ab8245, Abcam) [ ], V5 (catalog no. R96025, Invitrogen), pJNK (catalog no. 9255, Cell Signaling Technology) [ ] and JNK3 (catalog no. CP10162, Cell Applications) were used.

Techniques: Western Blot, Expressing, Activation Assay

Journal: Cell Death and Differentiation

Article Title: APP upregulation contributes to retinal ganglion cell degeneration via JNK3

doi: 10.1038/s41418-017-0005-3

Figure Lengend Snippet: a – d Representative western blots show JNK3 expression and overall phosphorylative activation of JNK in SH-SY5Y-APP cells treated with vehicle or γ-secretase inhibitors a L-685,458 (5 μM) or c BMS 299897 (1 μM). b , d The bar charts show quantitative analysis of relative JNK3 expression and overall phosphorylative activation of JNK normalized to the GAPDH ( n = 3 for each treatment). e Representative western blots show phosphorylative activation of JNK3 in retinae from vehicle-treated or γ-secretase inhibitor BMS 299897 treated mice 2 weeks after ONA. f Representative fluorescence microscope images show immunostaining of mouse retina flat-mounts from vehicle or γ-secretase inhibitor BMS 299897 treated mice 2 weeks after ONA using anti-Tuj1 antibody. Scale bar, 50 μm. g The bar chart shows quantitative analysis of the surviving Tuj1 + RGC number in injured mouse retinae 14 days after ONA ( n = 4 for each treatment). Student’s t -test: ** p < 0.01. Error bars represent SEM

Article Snippet: Rabbit antibodies against APP (Y188, Abcam) [ ], JNK1 (catalog no. MAB17761, R&D Systems), JNK2 (catalog no. MAB1846, R&D Systems) [ ], JNK3 (catalog no. 2305, Cell Signaling Technology), p-cJun73 (catalog no. 9164, Cell Signaling Technology) and c-Jun (catalog no. 9165, Cell Signaling Technology); mouse antibodies against GAPDH (catalog no. ab8245, Abcam) [ ], V5 (catalog no. R96025, Invitrogen), pJNK (catalog no. 9255, Cell Signaling Technology) [ ] and JNK3 (catalog no. CP10162, Cell Applications) were used.

Techniques: Western Blot, Expressing, Activation Assay, Fluorescence, Microscopy, Immunostaining

Journal: Cell Death and Differentiation

Article Title: APP upregulation contributes to retinal ganglion cell degeneration via JNK3

doi: 10.1038/s41418-017-0005-3

Figure Lengend Snippet: ONA induces upregulation of APP expression. APP is sequentially cleaved by α- or β-secretase and γ-secretase, producing several fragments and AICD, which is inhibited by APP deficiency or BMS 299897. AICD is translocated to the nucleus and upregulates JNK3 expression (probably functioning with other partners). Phosphorylated JNK3 then contributes to RGC death

Article Snippet: Rabbit antibodies against APP (Y188, Abcam) [ ], JNK1 (catalog no. MAB17761, R&D Systems), JNK2 (catalog no. MAB1846, R&D Systems) [ ], JNK3 (catalog no. 2305, Cell Signaling Technology), p-cJun73 (catalog no. 9164, Cell Signaling Technology) and c-Jun (catalog no. 9165, Cell Signaling Technology); mouse antibodies against GAPDH (catalog no. ab8245, Abcam) [ ], V5 (catalog no. R96025, Invitrogen), pJNK (catalog no. 9255, Cell Signaling Technology) [ ] and JNK3 (catalog no. CP10162, Cell Applications) were used.

Techniques: Expressing

Journal: Biochemical pharmacology

Article Title: The signalling axis mediating neuronal apoptosis in response to [Pt(O,O'-acac)(γ-acac)(DMS)].

doi: 10.1016/j.bcp.2011.03.007

Figure Lengend Snippet: Fig. 4. Effect of [Pt(O,O0-acac)(g-acac)(DMS)] on Dcm, apoptotic and signal transduction proteins in SH-SY5Y cells. (A) Fluorescent spectra of JC-1 in SH-SY5Y cells treated or not with 10 mM [Pt(O,O0-acac)(g-acac)(DMS)] for the indicated time. The data are means S.D. of three different experiments and are presented as red J-aggregates/green monomer JC-1 fluorescence ratio. Asterisks indicate values that are significantly different (p < 0.05) from control at the same concentration and time point. (B) Mitochondrial fractions were prepared at the indicated times of [Pt(O,O0-acac)(g-acac)(DMS)] treatment, and the kinetics of Bax and Bcl-2 cytosol-to-mitochondria translocations and the release of cytochrome c were examined. Porin served as a mitochondrial indicator. (C) Cells were treated or not with 10 mM [Pt(O,O0-acac)(g-acac)(DMS)] for indicated time. Cell lysates were analysed by Western blotting with anti-unphosphorylated and phosphorylated p38MAPK, ERK1/2, JNK1/2, and Akt antibodies. IP: the immunoprecipitated sample obtained by a specific anti total JNK3 monoclonal antibody was analysed by Western blotting using an anti-unphosphorylated (JNK3) and phosphorylated (pJNK3) antibodies. The same blots were stripped and reprobed with an anti-b-actin monoclonal antibody. Representative immunoblots of three experiments are depicted.

Article Snippet: Anti-porin (or anti-voltage-dependent anion selective channel 1, VDAC1), anti-p65 antibody, anti total (phosphorylated and unphosphorylated) JNK3, anti-histone-3/4 antibodies, goat antirabbit IgG conjugated with peroxidase, as well as control antibodies, were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Transduction, Control, Concentration Assay, Western Blot, Immunoprecipitation

Journal: Investigative ophthalmology & visual science

Article Title: Down-Regulation of RNA 3'-Terminal Phosphate Cyclase Attenuates Toll-Like Receptor 3-Mediated Axonal Loss in the Retina and Optic Nerve.

doi: 10.1167/iovs.16-19799

Figure Lengend Snippet: FIGURE 5. Rtca, JNK3, and pJNK3 levels were not up-regulated in Tlr3 knockout mice. At 48 hours after treating C57BL/6J and Tlr3 knockout mouse eyes with PBS or Poly(I:C), Western blot analysis was performed on total proteins extracted from the retinas by using Rtca, JNK3, and pJNK3 antibodies (A). Actin bands show the loading of an equal amount of total proteins. The bar graph in B shows semiquantitative analysis of Rtca, JNK3, and pJNK3 levels from the Western blots. *P < 0.02 when Rtca, JNK3, and pJNK3 levels in retinal proteins extracted from Poly(I:C)-treated C57BL/6J mouse eyes were compared with Rtca, JNK3, and pJNK3 in retinal proteins extracted from PBS-treated C57BL/6J mouse eyes. **P < 0.03 when Rtca, JNK3, and pJNK3 levels in retinal protein extracted from Poly(I:C)-treated Tlr3 knockout mice were compared with Poly(I:C)-treated C57BL/6J mouse eyes.

Article Snippet: The c-jun N-terminal kinase 3 (JNK3; 2305) and phospho-JNK3 (pJNK3) (9251) antibodies were obtained from Novus Biologicals (Littleton, CO, USA).

Techniques: Knock-Out, Western Blot

Journal: Investigative ophthalmology & visual science

Article Title: Down-Regulation of RNA 3'-Terminal Phosphate Cyclase Attenuates Toll-Like Receptor 3-Mediated Axonal Loss in the Retina and Optic Nerve.

doi: 10.1167/iovs.16-19799

Figure Lengend Snippet: FIGURE 7. Down-regulation of Rtca, JNK3, and pJNK3 in the retinas from Rtca siRNA-treated C57BL/6J mice. Equal amounts of retinal proteins (50 lg) extracted from the eyes treated with control RNA or Rtca siRNA, with or without PBS or Poly(I:C) were subjected to Western blot analysis by using antibodies for Rtca, JNK3, and pJNK3 (A). Actin bands indicate loading controls for the total proteins. The bar graph (B) shows the relative levels of Rtca, JNK3, and pJNK3 in retinal protein extracts. *P < 0.03 and #P < 0.04 when Rtca, JNK3, and pJNK3 levels in the retinal proteins extracted from Poly(I:C)-treated eyes were compared with Rtca, JNK3, and pJNK3 levels in the retinal proteins extracted from PBS, PBS plus Rtca siRNA, and PBS plus control RNA-treated eyes. **P < 0.02 when Rtca, JNK3, and pJNK3 levels in retinal proteins extracted from Poly(I:C) plus Rtca siRNA-treated eyes were compared with Rtca, JNK3, and pJNK3 levels in Poly(I:C) or Poly(I:C) plus control RNA-treated eyes.

Article Snippet: The c-jun N-terminal kinase 3 (JNK3; 2305) and phospho-JNK3 (pJNK3) (9251) antibodies were obtained from Novus Biologicals (Littleton, CO, USA).

Techniques: Control, Western Blot

Journal: Journal of neuroinflammation

Article Title: SPOCK2 modulates neuropathic pain by interacting with MT1-MMP to regulate astrocytic MMP-2 activation in rats with chronic constriction injury.

doi: 10.1186/s12974-024-03051-5

Figure Lengend Snippet: Fig. 5 RNAi-mediated SPOCK2 downregulation in CCI rats reduces astrocytic activation, ERK1/2 activation and IL-1β production. A Double staining showing decreased (**P < 0.01 vs CCI + NC-siRNA) colocalization of SPOCK2 with GFAP after i.t. injection of SPOCK2-siRNA (Scale 50 um). B Decreased expression ratio of p-ERK/ERK after i.t. injection of SPOCK2-siRNA. (**P < 0.01 vs CCI + NC-siRNA). The expression ratio changes of p-JNK/ JNK were not significant (P > 0.05 vs CCI + NC-siRNA) after i.t. injection of SPOCK2-siRNA. C Decreased expression level of IL-1β after i.t. injection of SPOCK2-siRNA. (**P < 0.01 vs CCI + NC-siRNA). The change in expression of CCL2 was not significant (P > 0.05 vs CCI + NC-siRNA) after i.t. injection of SPOCK2-siRNA

Article Snippet: The membranes were next placed in blocking buffer (5% milk in Tris-buffered saline with Tween-20) for 1 h and incubated overnight (16–18 h) at 4 °C with primary antibodies against SPOCK2 (1:1,000, Cat# 217,044, RRID: AB_3076329, Abcam, Cambridge, UK), MT1-MMP (1:1,000, Cat# 14,552–1-AP, RRID: AB_2250751, Proteintech, Wuhan, China), MMP-2 (1:1,000, Cat# 66,366–1-Ig, RRID: AB_2881746, Proteintech), ERK1/2 (1:2,000, Cat# 9102, RRID: AB_330744, Cell Signaling Technology, Danvers, MA, USA), p-ERK1/2 (1:1,000, Cat# 4370, RRID: AB_2315112, Cell Signaling Technology), interleukin-1β (IL-1β) (1:1,000, Cat# sc-7884, RRID: AB_2124476, Santa Cruz, TX, USA), CC-chemokine ligand 2 (CCL2) (1:2000, Cat# ab7202, RRID: AB_305755, Abcam), JNK (1:3,000, Cat# 17,572–1-AP, RRID: AB_2266214, Proteintech), p-JNK (1:2,000, Cat# 80,024–1-RR, RRID: AB_2882943, Proteintech), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (1:10,000, Cat# 60,004–1-Ig, RRID: AB_2107436, Proteintech).

Techniques: Activation Assay, Double Staining, Injection, Expressing

Journal: Poultry Science

Article Title: Effect of heat stress on mitogen-activated protein kinases in the hypothalamic–pituitary–gonadal axis of developing Wenchang chicks 1

doi: 10.3382/ps/pez499

Figure Lengend Snippet: Effect of heat stress on phospho-MAPKs in the hypothalamus of chicks (× 400, Bar = 50 μm, MIOD/μm 2 ). Group A and group B are female and male chicks, respectively, followed by group C and group D. Control check (CK); heat stress (HS); and the numbers after CK/HS detail weeks from 1 to 6; phospho-ERK (pERK); phospho-JNK (pJNK); phospho-P38 (pP38); mean integral optical density (MIOD). Positive reactions are denoted with an arrow. * indicates P < 0.05 and ** indicates P < 0.01 between the CK group and the HS group in the same week, and the same below.

Article Snippet: After incubating with primary antibodies of rabbit phospho-ERK (Cat# bs-1646R), phospho-JNK (Cat# bs-17591R), or phospho-p38 (Cat# bs-2210R) (Bioss Biotech, Beijing, China) at 1:400 dilution, the sections were further incubated with the secondary antibody (Yeasen Biotech Co. Ltd., Shanghai, China).

Techniques:

Journal: Poultry Science

Article Title: Effect of heat stress on mitogen-activated protein kinases in the hypothalamic–pituitary–gonadal axis of developing Wenchang chicks 1

doi: 10.3382/ps/pez499

Figure Lengend Snippet: Effect of heat stress on phospho-/total MAPKs in the hypothalamus of chicks. Group A is the histogram result of the Western Blot followed by group B. Control check (CK); heat stress (HS); phospho-ERK (pERK); phospho-JNK (pJNK); phospho-P38 (pP38); total ERK (tERK); total JNK (tJNK); total P38 (tP38). * indicates P < 0.05 and ** indicates P < 0.01 between the CK group and the HS group in the same week, and the same below.

Article Snippet: After incubating with primary antibodies of rabbit phospho-ERK (Cat# bs-1646R), phospho-JNK (Cat# bs-17591R), or phospho-p38 (Cat# bs-2210R) (Bioss Biotech, Beijing, China) at 1:400 dilution, the sections were further incubated with the secondary antibody (Yeasen Biotech Co. Ltd., Shanghai, China).

Techniques: Western Blot

Journal: Poultry Science

Article Title: Effect of heat stress on mitogen-activated protein kinases in the hypothalamic–pituitary–gonadal axis of developing Wenchang chicks 1

doi: 10.3382/ps/pez499

Figure Lengend Snippet: Effect of crosstalk among MAPKs signaling pathways in the hypothalamus of chicks. ♀ and ♂ reflect female and male chicks, respectively. phospho-ERK (pERK); phospho-JNK (pJNK); phospho-P38 (pP38), and the same below.

Article Snippet: After incubating with primary antibodies of rabbit phospho-ERK (Cat# bs-1646R), phospho-JNK (Cat# bs-17591R), or phospho-p38 (Cat# bs-2210R) (Bioss Biotech, Beijing, China) at 1:400 dilution, the sections were further incubated with the secondary antibody (Yeasen Biotech Co. Ltd., Shanghai, China).

Techniques:

Journal: Behavioral and Brain Functions : BBF

Article Title: Astragalus injection protects cerebral ischemic injury by inhibiting neuronal apoptosis and the expression of JNK3 after cerebral ischemia reperfusion in rats

doi: 10.1186/1744-9081-9-36

Figure Lengend Snippet: The expression of JNK3 in parahipppocampal area, SP staining × 400. (A) Control group: neurons arranged neatly, cell structure was completely. (B) Model group: neurons arranged disorderly, the number of JNK3-positive neurons increased significantly. (C) Treatment group: the number of JNK3-positive neurons (→) decreased significantly.

Article Snippet: The rabbit anti-rat JNK3 antibody and immunohistochemical SP kit (Wuhan Boster Biotech.

Techniques: Expressing, Staining, Control

Journal: Behavioral and Brain Functions : BBF

Article Title: Astragalus injection protects cerebral ischemic injury by inhibiting neuronal apoptosis and the expression of JNK3 after cerebral ischemia reperfusion in rats

doi: 10.1186/1744-9081-9-36

Figure Lengend Snippet: The fragmentation formed by JNK3 protein detected by Western blotting. Line 1: a weaker fragment was formed by JNK3 protein in control group. Line 2: a strong fragment was formed by JNK3 protein in model group. Line 3: a middle fragment was formed by JNK3 protein in treatment group.

Article Snippet: The rabbit anti-rat JNK3 antibody and immunohistochemical SP kit (Wuhan Boster Biotech.

Techniques: Western Blot, Control

Journal: Behavioral and Brain Functions : BBF

Article Title: Astragalus injection protects cerebral ischemic injury by inhibiting neuronal apoptosis and the expression of JNK3 after cerebral ischemia reperfusion in rats

doi: 10.1186/1744-9081-9-36

Figure Lengend Snippet: Electrophoresis analysis of RT-PCR products of JNK3 mRNA (left) and GAPDH (right). M: Maker; Line 1–2: weak light bands in control group; Line 3–4: strong light bands in model group; Line 5–6: middle light bands in treatment group

Article Snippet: The rabbit anti-rat JNK3 antibody and immunohistochemical SP kit (Wuhan Boster Biotech.

Techniques: Electrophoresis, Reverse Transcription Polymerase Chain Reaction, Control

Journal: Frontiers in Veterinary Science

Article Title: Adiponectin receptor agonist reduces broiler hepatic lipid deposition

doi: 10.3389/fvets.2025.1667501

Figure Lengend Snippet: Effects of AdipoRon on lipid accumulation, AMPK/PPARα and JNK1 signaling pathway in the liver of CORT broilers. (A) Immunoblot of ACC, CPT-1, PPARα and ADPN protein level in the liver of CORT broiler. (B) The change of ACC, CPT-1, PPARα and ADPN protein expression in the liver of CORT broiler. (C) Immunoblot of p-AMPKα1, AMPKα1, p-JNK1, JNK1 and TNFα protein level in the liver of CORT broiler. (D) The change of p-AMPKα1, AMPKα1, p-JNK1, JNK1 and TNFα protein expression in the liver of CORT broiler. Grayscale values of each band were analyzed using ImageJ software. Normalization was performed by separately comparing the grayscale values of target protein bands with those of corresponding loading control bands (GAPDH), as well as the grayscale values of phosphorylated protein bands with those of total protein bands. The data represent mean ± SEM. Differences were determined by one-way ANOVA followed by Tukey’s test The bars with different small letter differ significantly between groups ( p < 0.05, n = 6, biological replicates per group). ACC, Acetyl-CoA carboxylase 1; CPT-1, carnitine palmitoyl transferase-1; PPARα, peroxisome proliferators-activated receptor α; ADPN, adiponectin; AMPKα1, adenosine 5′-monophosphate (AMP)-activated protein kinase alpha 1; p-AMPKα1, phosphorylated adenosine 5′-monophosphate (AMP)-activated protein kinase alpha 1; JNK1, c-Jun N-terminal kinase 1; p-JNK1, phosphorylated c-Jun N-terminal kinase 1; TNF-α, Tumor Necrosis Factor-alpha.

Article Snippet: The membranes were blocked with a 5% skim milk powder solution at room temperature for 2 h. Subsequently, membranes were incubated with primary antibodies at 4°C for 12 h. After incubation with corresponding secondary antibodies at RT for 1 h. The primary antibodies used in this study included: ADPN (bs-0471R; Bioss, Beijing, China), PPAR α (bs-3614R; Bioss, Beijing, China), TNF-α (bsm-33207 M; Bioss, Beijing, China), p-AMPK (bs-5551R; Bioss, Beijing, China), AMPK (bs-41337R; Bioss, Beijing, China), p-JNK1 (bs-17591R; Bioss, Beijing, China), JNK1 (bs-20760R; Bioss, Beijing, China), and GAPDH (bsm-33033 M; Bioss, Beijing, China) as an internal control.

Techniques: Western Blot, Expressing, Software, Control

Journal: Frontiers in Veterinary Science

Article Title: Adiponectin receptor agonist reduces broiler hepatic lipid deposition

doi: 10.3389/fvets.2025.1667501

Figure Lengend Snippet: Effects of AdipoRon on viability, lipid accumulation, AMPK/PPARα and JNK1 signaling pathway in FE-induced LMH cells. (A) The cell viability. (B) The change of oil red O quantification in LMH cells. (C1–3) Liver oil red O staining in LMH cells. Bar = 20 μm. (D) Immunoblot of ACC, CPT-1, PPARα, ADPN, p-AMPKα1, AMPKα1 PPARα, p-JNK1, JNK1 and TNF-α protein level in LMH cells. (E) The change of ACC, CPT-1, PPARα, ADPN, p-AMPKα1, AMPKα1, PPARα, p-JNK1, JNK1 and TNF-α protein expression in LMH cells. Grayscale values of each band were analyzed using ImageJ software. Normalization was performed by separately comparing the grayscale values of target protein bands with those of corresponding loading control bands (GAPDH), as well as the grayscale values of phosphorylated protein bands with those of total protein bands. The data represent mean ± SEM. Differences were determined by one-way ANOVA followed by Tukey’s test The bars with different small letter (a, b, c) differ significantly between groups ( p < 0.05, n = 6 per group, biological replicates). FE, fat emulsion; AdipoRon, adiponectin receptor agonists; ACC, Acetyl-CoA carboxylase 1; CPT-1, carnitine palmitoyl transferase-1; PPARα, peroxisome proliferators-activated receptor α; ADPN, adiponectin; TNF-α, tumor necrosis factor-alpha; AMPKα1, adenosine 5′-monophosphate (AMP)-activated protein kinase alpha 1; p-AMPKα1, phosphorylated adenosine 5′-monophosphate (AMP)-activated protein kinase alpha 1; JNK1, c-Jun N-terminal kinase 1; p-JNK1, phosphorylated c-Jun N-terminal kinase 1.

Article Snippet: The membranes were blocked with a 5% skim milk powder solution at room temperature for 2 h. Subsequently, membranes were incubated with primary antibodies at 4°C for 12 h. After incubation with corresponding secondary antibodies at RT for 1 h. The primary antibodies used in this study included: ADPN (bs-0471R; Bioss, Beijing, China), PPAR α (bs-3614R; Bioss, Beijing, China), TNF-α (bsm-33207 M; Bioss, Beijing, China), p-AMPK (bs-5551R; Bioss, Beijing, China), AMPK (bs-41337R; Bioss, Beijing, China), p-JNK1 (bs-17591R; Bioss, Beijing, China), JNK1 (bs-20760R; Bioss, Beijing, China), and GAPDH (bsm-33033 M; Bioss, Beijing, China) as an internal control.

Techniques: Staining, Western Blot, Expressing, Software, Control, Emulsion

Journal: International Journal of Molecular Sciences

Article Title: Preparation of CPD Photolyase Nanoliposomes Derived from Antarctic Microalgae and Their Effect on UVB-Induced Skin Damage in Mice

doi: 10.3390/ijms232315148

Figure Lengend Snippet: The effect of CPDNL on mRNA expression of MAPK/AP-1/MMP signaling pathways in UVB-induced skin. ( a–c ) The mRNA expression of ERK, JNK, and P38; ( d ) the mRNA expression of AP-1; ( e , f ) the mRNA expression of MMP-1 and MMP-2. Data are presented as mean ± SEM. ** p < 0.01 and *** p < 0.001.

Article Snippet: The membrane was incubated with the primary antibody against NF-κВ (BA1297-2, Boster, Wuhan, China), TNF-α (BA14901, Boster), IL-6 (BA4339-2, Boster), COX-2 (BA3708, Boster), ERK (GB112238, Servicebio, Wuhan, China), JNK (M02608-3, Boster), P38 (A00176-2, Boster), AP-1 (GB11270, Servicebio), MMP-1 (A00733-1, Boster), and MMP-2 (A00286, Boster) at 4 °C overnight, followed by washing at TBST, and then incubated with goat anti-rabbit HRP (horseradish peroxidase)-IgG (BA1055, Boster) as the secondary antibody for 45 min. After washed with TBST 3 times for 10 min, membrane was incubated in SuperSignal ECL Western substrate (Biosharp, Hefei, China).

Techniques: Expressing, Protein-Protein interactions

Journal: International Journal of Molecular Sciences

Article Title: Preparation of CPD Photolyase Nanoliposomes Derived from Antarctic Microalgae and Their Effect on UVB-Induced Skin Damage in Mice

doi: 10.3390/ijms232315148

Figure Lengend Snippet: The effect of CPDNL on protein level of MAPK/AP-1/MMP signaling pathways in UVB-induced skin. The protein expression level ( a ) of ERK ( b ), JNK ( c ), P38 ( d ), AP-1 ( e ), MMP-1 ( f ), and MMP-2 ( g ) was determined by Western blot analysis. Data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: The membrane was incubated with the primary antibody against NF-κВ (BA1297-2, Boster, Wuhan, China), TNF-α (BA14901, Boster), IL-6 (BA4339-2, Boster), COX-2 (BA3708, Boster), ERK (GB112238, Servicebio, Wuhan, China), JNK (M02608-3, Boster), P38 (A00176-2, Boster), AP-1 (GB11270, Servicebio), MMP-1 (A00733-1, Boster), and MMP-2 (A00286, Boster) at 4 °C overnight, followed by washing at TBST, and then incubated with goat anti-rabbit HRP (horseradish peroxidase)-IgG (BA1055, Boster) as the secondary antibody for 45 min. After washed with TBST 3 times for 10 min, membrane was incubated in SuperSignal ECL Western substrate (Biosharp, Hefei, China).

Techniques: Protein-Protein interactions, Expressing, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Preparation of CPD Photolyase Nanoliposomes Derived from Antarctic Microalgae and Their Effect on UVB-Induced Skin Damage in Mice

doi: 10.3390/ijms232315148

Figure Lengend Snippet: Real-time quantitative PCR primer sequences.

Article Snippet: The membrane was incubated with the primary antibody against NF-κВ (BA1297-2, Boster, Wuhan, China), TNF-α (BA14901, Boster), IL-6 (BA4339-2, Boster), COX-2 (BA3708, Boster), ERK (GB112238, Servicebio, Wuhan, China), JNK (M02608-3, Boster), P38 (A00176-2, Boster), AP-1 (GB11270, Servicebio), MMP-1 (A00733-1, Boster), and MMP-2 (A00286, Boster) at 4 °C overnight, followed by washing at TBST, and then incubated with goat anti-rabbit HRP (horseradish peroxidase)-IgG (BA1055, Boster) as the secondary antibody for 45 min. After washed with TBST 3 times for 10 min, membrane was incubated in SuperSignal ECL Western substrate (Biosharp, Hefei, China).

Techniques: Real-time Polymerase Chain Reaction