mapk signaling pathway activation  (Galectin Therapeutics)

Journal: PLOS One

Article Title: Proteomic profiling of small extracellular vesicles from bovine nucleus pulposus cells

doi: 10.1371/journal.pone.0324179

Figure Lengend Snippet: A) STRING network of 141 more abundant NP small EV proteins; B) Cluster 1: Complement component C1q complex (red), complement and coagulation cascades (blue), complement activation classical pathway (green); C) Cluster 2: Proteasome (red); D) Cluster 3: Axon (blue), axon guidance receptor activity (red), Ephrin signaling (green); E) Cluster 4: Ras signaling (red), axon guidance (green) and MAPK signaling pathway (blue); F) Cluster 5: Hemostasis (green), regulation of MAPK cascade (blue), complement and coagulation cascades (red); G) Cluster 6: ECM (pink), integrin binding (light green), collagen binding (sky blue), ECM receptor interaction (red), PI3K/AKT signaling pathway (blue), ECM organization (yellow), signaling by RTKs (dark green). STRING: Search tool for the retrieval of interacting genes/proteins; NP: Nucleus pulposus.

Article Snippet: Environmental Information Processing , Regulation of actin cytoskeleton Focal adhesion Rap1 signaling pathway ECM-receptor interaction PI3K-Akt signaling pathway Chemokine signaling pathway Apelin signaling pathway Sphingolipid signaling pathway Ras signaling pathway cAMP signaling pathway MAPK signaling pathway , bta04810 bta04510 bta04015 bta04512 bta04151 bta04062 bta04371 bta04071 bta04014 bta04024 bta04010 , ACTN1, C9, COL6A1, COL6A3, FLNA, FN1, GNAI1, GNAI2, GNAI3, GNAQ, GNB1, GSN, HSPA8, HSPB1, HSPG2, ITGA3, ITGAV, ITGB1, LAMC1, MSN, MYH9, PFN1, RAB5B, RAB5C, RAC1, RHOA, RRAS, RRAS2, RAP1B, TLN1, YWHAQ, YWHAE, YWHAG, YWHAZ.

Techniques: Coagulation, Activation Assay, Activity Assay, Binding Assay

Journal: PLOS One

Article Title: Proteomic profiling of small extracellular vesicles from bovine nucleus pulposus cells

doi: 10.1371/journal.pone.0324179

Figure Lengend Snippet: A) STRING network of 484 NP small EV proteins; B) Cluster 1: Extracellular vesicle biogenesis (red), multivesicular body sorting pathway (blue), ESCRT I complex (pink), ESCRT (green), Flotillin complex (yellow); C) Cluster 2: Detection of oxidative stress (black), pentose phosphate pathway (red), glycolysis/gluconeogenesis (blue), biosynthesis of amino acids (green), carbon metabolism (pink), pyruvate metabolism (dark green), TCA cycle (yellow), glutathione metabolism (sky blue), HIF1 signaling pathway (purple), amino sugar and nucleotide sugar metabolism (ochre yellow), activation of BAD and translocation to mitochondria (brown), regulation of localization of FOXO transcription factors (grey); D) Cluster 3: Regulation of Schwann cell migration (blue), G-protein coupled receptor signaling pathway (green), axon guidance (red); E) Cluster 4: Rac protein signal transduction (red), RAS protein signal transduction (blue), small GTPase mediated signal transduction (green), RAP1 signaling pathway (yellow), EPHB-mediated forward signaling (pink), VEGFA/VEGFR2 pathway (dark green), signaling by Rho GTPases (sky blue); F); Cluster 5: MAP2K and MAPK activation (red), RAS signaling pathway (green), MAPK signaling pathway (pink), PI3K/AKT signaling pathway (yellow); G) Cluster 6 version 1: Cell adhesion mediated by integrin (red), mesodermal cell differentiation (blue), angiogenesis (green), regulation of small GTPase mediated signal transduction (yellow), negative regulation of apoptotic process (pink); H) Cluster 6 version 2: ECM receptor interaction (red), focal adhesion (green), PI3K/AKT signaling pathway (blue), axon guidance (yellow), TGFβ signaling pathway (pink); I) Cluster 7: Positive regulation of lamellipodium assembly (blue), actin cytoskeleton organization (red), EPHB mediated forward signaling (green); J) Cluster 8: ECM assembly (pink), cartilage development (green), angiogenesis (blue), blood vessel development (red), cell adhesion (yellow), PI3K/AKT signaling pathway (dark green). ECM: Extracellular matrix; ESCRT I: Endosomal sorting complex required for transport I; STRING: Search tool for the retrieval of interacting genes/proteins; NP: Nucleus pulposus.

Article Snippet: Environmental Information Processing , Regulation of actin cytoskeleton Focal adhesion Rap1 signaling pathway ECM-receptor interaction PI3K-Akt signaling pathway Chemokine signaling pathway Apelin signaling pathway Sphingolipid signaling pathway Ras signaling pathway cAMP signaling pathway MAPK signaling pathway , bta04810 bta04510 bta04015 bta04512 bta04151 bta04062 bta04371 bta04071 bta04014 bta04024 bta04010 , ACTN1, C9, COL6A1, COL6A3, FLNA, FN1, GNAI1, GNAI2, GNAI3, GNAQ, GNB1, GSN, HSPA8, HSPB1, HSPG2, ITGA3, ITGAV, ITGB1, LAMC1, MSN, MYH9, PFN1, RAB5B, RAB5C, RAC1, RHOA, RRAS, RRAS2, RAP1B, TLN1, YWHAQ, YWHAE, YWHAG, YWHAZ.

Techniques: Activation Assay, Translocation Assay, Migration, Transduction, Cell Differentiation

Journal: PLOS One

Article Title: Proteomic profiling of small extracellular vesicles from bovine nucleus pulposus cells

doi: 10.1371/journal.pone.0324179

Figure Lengend Snippet: NP small EV protein cargo and membrane constituents are involved in key metabolic pathways, including glycolysis, gluconeogenesis, Krebs cycle and PPP. They are crucial for energy production and to maintain the cellular redox balance. NP small EVs with the aid of the 20S proteasome, ensure protein quality control and reduced inflammation in a recipient cell. NP small EVs modulate signaling through EPH receptors, impacting cellular communication and tissue organization. Additionally, NP small EV interact with the complement system, influencing the classical, alternative, and lectin pathways involved in immune responses and inflammation. The PI3K/AKT/RAS signaling pathway is axis is impacted by the NP small EV proteome, which could promote ECM synthesis, cell growth and proliferation. Together, these processes underscore the essential role of the NP small EV proteome in sustaining NP cell function and NP niche homeostasis. ECM: Extracellular matrix; EPH: Ephrin receptor, ER: endoplasmic reticulum, ERK: extracellular signal-regulated kinase, MEK: mitogen-activated protein kinase, NP: nucleus pulposus, PDK1: phosphoinositide-dependent protein kinase 1, PPP: pentose phosphate pathway, PI3K/AKT: phosphoinositide 3-kinase/protein kinase B, RAF: rapidly accelerated fibrosarcoma, RHEB: RAS homolog enriched in brain, mTORC1: mammalian target of rapamycin complex 1, small EV: small extracellular vesicles, TSC1/2: tuberous sclerosis proteins 1 and 2. This illustration was created on Biorender. ( www.biorender.com/ ).

Article Snippet: Environmental Information Processing , Regulation of actin cytoskeleton Focal adhesion Rap1 signaling pathway ECM-receptor interaction PI3K-Akt signaling pathway Chemokine signaling pathway Apelin signaling pathway Sphingolipid signaling pathway Ras signaling pathway cAMP signaling pathway MAPK signaling pathway , bta04810 bta04510 bta04015 bta04512 bta04151 bta04062 bta04371 bta04071 bta04014 bta04024 bta04010 , ACTN1, C9, COL6A1, COL6A3, FLNA, FN1, GNAI1, GNAI2, GNAI3, GNAQ, GNB1, GSN, HSPA8, HSPB1, HSPG2, ITGA3, ITGAV, ITGB1, LAMC1, MSN, MYH9, PFN1, RAB5B, RAB5C, RAC1, RHOA, RRAS, RRAS2, RAP1B, TLN1, YWHAQ, YWHAE, YWHAG, YWHAZ.

Techniques: Membrane, Control, Cell Function Assay

Journal: Frontiers in Pharmacology

Article Title: Glycyrrhetinic Acid Protects Renal Tubular Cells against Oxidative Injury via Reciprocal Regulation of JNK-Connexin 43-Thioredoxin 1 Signaling

doi: 10.3389/fphar.2021.619567

Figure Lengend Snippet: c-Jun N-terminal kinase (JNK)-mediated oxidative stress induces connexin 43 (Cx43). (A) Induction of JNK phosphorylation and expression of Cx43 by Px-12. NRK-52E cells in 6-well plates were incubated with Px-12 for 0, 1, 3, 6, 9, and 12 h separately. Cellular lysates were then analyzed by western blotting for phosphorylated JNK and Cx43 expression. Densitometric analysis of p -JNK and Cx43 are shown on the right (mean ± SD, n = 3; ** p < 0.01 vs. the control, ## p < 0.01 vs. Px-12 in control). (B) Effects of 18α-glycyrrhetinic acid (Ga) and sp600125, an inhibitor of JNK, on Px12-induced cell injury. NRK-52E cells were pretreated with Ga (20 μM) and sp600125 (20 μM) for 1 h and then challenged with Px-12 for another 24 h. Then, cell viability was evaluated using a Cell Counting Kit-8 (CCK-8) assay. Data are expressed as the percentage of living cells vs. the untreated control (mean ± SD, n = 6). * p < 0.01 vs. the control. ## p < 0.01 vs. Px-12 in control. (C) Influence of sp600125 and Ga on NRK-52E cell apoptosis staining. NRK-52E cells in 96-well plate were pretreated with 18α-Ga and sp600125 for 1 h and then incubated with Px-12 for another 24 h. Then, apoptotic cells were stained via DT-mediated dUTP nick end labeling (TUNEL) and 4,6-diamidino-2-phenylindole (DAPI) (magnification, ×400). The ratio of positive cells is shown on the right (mean ± SD, n = 3; ** p < 0.01 vs. the control, ## p < 0.01 vs. Px-12 in the control).

Article Snippet: The JNK-mitogen-activated protein kinase (MAPK) signaling pathway inhibitor (sp600125, A4604) was obtained from ApexBio Technology China.

Techniques: Expressing, Incubation, Western Blot, Cell Counting, CCK-8 Assay, Staining, End Labeling, TUNEL Assay

Journal: Frontiers in Pharmacology

Article Title: Glycyrrhetinic Acid Protects Renal Tubular Cells against Oxidative Injury via Reciprocal Regulation of JNK-Connexin 43-Thioredoxin 1 Signaling

doi: 10.3389/fphar.2021.619567

Figure Lengend Snippet: Reciprocal modulation of connexin 43 (Cx43) and c-Jun N-terminal kinase (JNK). (A) Effect of 18α-glycyrrhetinic acid (Ga) and sp600125 on JNK phosphorylation and expression of Cx43. NRK-52E cells in six-well plates were pretreated with Ga and sp600125 for 1 h and then challenged with Px-12 for another 1 h. Cellular lysates were then analyzed by western blotting for phosphorylated JNK and Cx43 expression. Densitometric analysis of p -JNK and Cx43 are shown on the right (mean ± SD, n = 3; ** p < 0.01 vs. the control, ## p < 0.01 vs. Px-12 in the control). (B) NRK-52E cells were transfected with either Cx43 siRNA or siRNA control for 24 h. Cellular lysates were analyzed by western blotting for Cx43 expression. Densitometric analysis of Cx43 is shown on the right (mean ± SD, n = 3; ** p < 0.01 vs. the siRNA control group). (C) Effect of Cx43 siRNA on JNK phosphorylation and Cx43 expression. The transfected or normal cells were incubated with Px-12 for 1 h separately. Then, cellular lysates were analyzed by western blotting for phosphorylated JNK and Cx43 expression. Densitometric analysis of p -JNK and Cx43 are shown on the right (mean ± SD, n = 3; ** p < 0.01 vs. the control, ## p < 0.01 vs. Px-12 in the siRNA control group). (D) Effect of Cx43 siRNA on cell viability. The transfected or normal cells were incubated with Px-12 for 24 h. Then, cell viability was evaluated using a Cell Counting Kit-8 (CCK-8) assay. Data are expressed as the percentage of living cells vs. the untreated control (mean ± SD, n = 6). * p < 0.01 vs. Px-12 in normal cells. ## p < 0.01 vs. Px12 in the siRNA control group. (E) Effect of Cx43 siRNA on caspase-3 activation. The transfected or normal cells were incubated with Px-12 for 24 h. Then, cellular lysates were analyzed by western blotting for expression of cleaved caspase-3. Densitometric analysis of cleaved caspase-3 and caspase-3 are shown on the right (mean ± SD, n = 3; ** p < 0.01 vs. Px-12 in the siRNA control group).

Article Snippet: The JNK-mitogen-activated protein kinase (MAPK) signaling pathway inhibitor (sp600125, A4604) was obtained from ApexBio Technology China.

Techniques: Expressing, Western Blot, Transfection, Incubation, Cell Counting, CCK-8 Assay, Activation Assay

Journal: Frontiers in Pharmacology

Article Title: Glycyrrhetinic Acid Protects Renal Tubular Cells against Oxidative Injury via Reciprocal Regulation of JNK-Connexin 43-Thioredoxin 1 Signaling

doi: 10.3389/fphar.2021.619567

Figure Lengend Snippet: c-Jun N-terminal kinase (JNK)-mediated oxidative stress induces connexin 43 (Cx43). (A) Induction of JNK phosphorylation and expression of Cx43 by Px-12. NRK-52E cells in 6-well plates were incubated with Px-12 for 0, 1, 3, 6, 9, and 12 h separately. Cellular lysates were then analyzed by western blotting for phosphorylated JNK and Cx43 expression. Densitometric analysis of p -JNK and Cx43 are shown on the right (mean ± SD, n = 3; ** p < 0.01 vs. the control, ## p < 0.01 vs. Px-12 in control). (B) Effects of 18α-glycyrrhetinic acid (Ga) and sp600125, an inhibitor of JNK, on Px12-induced cell injury. NRK-52E cells were pretreated with Ga (20 μM) and sp600125 (20 μM) for 1 h and then challenged with Px-12 for another 24 h. Then, cell viability was evaluated using a Cell Counting Kit-8 (CCK-8) assay. Data are expressed as the percentage of living cells vs. the untreated control (mean ± SD, n = 6). * p < 0.01 vs. the control. ## p < 0.01 vs. Px-12 in control. (C) Influence of sp600125 and Ga on NRK-52E cell apoptosis staining. NRK-52E cells in 96-well plate were pretreated with 18α-Ga and sp600125 for 1 h and then incubated with Px-12 for another 24 h. Then, apoptotic cells were stained via DT-mediated dUTP nick end labeling (TUNEL) and 4,6-diamidino-2-phenylindole (DAPI) (magnification, ×400). The ratio of positive cells is shown on the right (mean ± SD, n = 3; ** p < 0.01 vs. the control, ## p < 0.01 vs. Px-12 in the control).

Article Snippet: The JNK-mitogen-activated protein kinase (MAPK) signaling pathway inhibitor (sp600125, A4604) was obtained from ApexBio Technology China.

Techniques: Expressing, Incubation, Western Blot, Cell Counting, CCK-8 Assay, Staining, End Labeling, TUNEL Assay

Journal: Frontiers in Pharmacology

Article Title: Glycyrrhetinic Acid Protects Renal Tubular Cells against Oxidative Injury via Reciprocal Regulation of JNK-Connexin 43-Thioredoxin 1 Signaling

doi: 10.3389/fphar.2021.619567

Figure Lengend Snippet: Reciprocal modulation of connexin 43 (Cx43) and c-Jun N-terminal kinase (JNK). (A) Effect of 18α-glycyrrhetinic acid (Ga) and sp600125 on JNK phosphorylation and expression of Cx43. NRK-52E cells in six-well plates were pretreated with Ga and sp600125 for 1 h and then challenged with Px-12 for another 1 h. Cellular lysates were then analyzed by western blotting for phosphorylated JNK and Cx43 expression. Densitometric analysis of p -JNK and Cx43 are shown on the right (mean ± SD, n = 3; ** p < 0.01 vs. the control, ## p < 0.01 vs. Px-12 in the control). (B) NRK-52E cells were transfected with either Cx43 siRNA or siRNA control for 24 h. Cellular lysates were analyzed by western blotting for Cx43 expression. Densitometric analysis of Cx43 is shown on the right (mean ± SD, n = 3; ** p < 0.01 vs. the siRNA control group). (C) Effect of Cx43 siRNA on JNK phosphorylation and Cx43 expression. The transfected or normal cells were incubated with Px-12 for 1 h separately. Then, cellular lysates were analyzed by western blotting for phosphorylated JNK and Cx43 expression. Densitometric analysis of p -JNK and Cx43 are shown on the right (mean ± SD, n = 3; ** p < 0.01 vs. the control, ## p < 0.01 vs. Px-12 in the siRNA control group). (D) Effect of Cx43 siRNA on cell viability. The transfected or normal cells were incubated with Px-12 for 24 h. Then, cell viability was evaluated using a Cell Counting Kit-8 (CCK-8) assay. Data are expressed as the percentage of living cells vs. the untreated control (mean ± SD, n = 6). * p < 0.01 vs. Px-12 in normal cells. ## p < 0.01 vs. Px12 in the siRNA control group. (E) Effect of Cx43 siRNA on caspase-3 activation. The transfected or normal cells were incubated with Px-12 for 24 h. Then, cellular lysates were analyzed by western blotting for expression of cleaved caspase-3. Densitometric analysis of cleaved caspase-3 and caspase-3 are shown on the right (mean ± SD, n = 3; ** p < 0.01 vs. Px-12 in the siRNA control group).

Article Snippet: The JNK-mitogen-activated protein kinase (MAPK) signaling pathway inhibitor (sp600125, A4604) was obtained from ApexBio Technology China.

Techniques: Expressing, Western Blot, Transfection, Incubation, Cell Counting, CCK-8 Assay, Activation Assay

Journal: Frontiers in Pharmacology

Article Title: Glycyrrhetinic Acid Protects Renal Tubular Cells against Oxidative Injury via Reciprocal Regulation of JNK-Connexin 43-Thioredoxin 1 Signaling

doi: 10.3389/fphar.2021.619567

Figure Lengend Snippet: Connexin 43 (Cx43) inhibition suppresses oxidative stress by regulating thioredoxin 1. (A) Effect of 18α-glycyrrhetinic acid (Ga) and sp600125 on reactive oxygens species (ROS)/O 2 − production. NRK-52E cells in a 96-well plate were pretreated with Ga and sp600125 separately for 1 h, challenged with Px-12 for another 1 h, and then incubated with a ROS/O 2 − probe for 30 min. Images were captured using an inverted fluorescence microscope (×100). Mean fluorescence intensity is shown at the bottom (mean ± SD, n = 3; ** p < 0.01 vs. the control, ## p < 0.01 vs. Px-12 in the control). (B) Effects of Ga on thioredoxin 1. NRK-52E cells in six-well plates were pretreated with Ga for 1 h and then challenged with Px-12 for another 1 h. Then, cellular lysates were analyzed by western blotting for the expression of thioredoxin 1. Densitometric analysis of thioredoxin 1, as determined by ImageJ, is shown on the right (mean ± SD, n = 3; ** p < 0.01 vs. the control, ## p < 0.01 vs. Px-12 in the siRNA control group).

Article Snippet: The JNK-mitogen-activated protein kinase (MAPK) signaling pathway inhibitor (sp600125, A4604) was obtained from ApexBio Technology China.

Techniques: Inhibition, Incubation, Fluorescence, Microscopy, Western Blot, Expressing