pkn2 mab  (R&D Systems)

Journal: Cancers

Article Title: Changes of Protein Expression after CRISPR/Cas9 Knockout of miRNA-142 in Cell Lines Derived from Diffuse Large B-Cell Lymphoma.

doi: 10.3390/cancers14205031

Figure Lengend Snippet: Figure 11. Western blot analysis: Extracts of BJAB-wild-type and SUDHL4-wild-type (“WT”) cells and the corresponding miR-142 knockout (“KO”) cells were separated on a 10% polyacrylamide gel, transferred to a nitrocellulose membrane and incubated with antibodies against PKN2, Ezrin and β-actin as a loading control. The membranes were incubated with the appropriate secondary antibodies coupled to horseradish peroxidase. Bound secondary antibodies were visualized by ECL. The bands were quantified using Image Lab 6.0.1. The mean value of two separate experiments is shown above the bands. The wild-type value was set to 1. The uncropped blots are shown in Figure S4.

Article Snippet: PKN2 antibody (Cat. Nr. 8697) was purchased from Cell Signaling (Frankfurt/M., Germany); a rat monoclonal antibody (rat IgG2a) directed against Ezrin was described elsewhere (T. Pfitzner, MD thesis, Saarland University Medical School, 66421 Homburg/Saar, Germany, 1996).

Techniques: Western Blot, Knock-Out, Membrane, Incubation, Control

Journal: American Journal of Physiology - Endocrinology and Metabolism

Article Title: Protein kinase N2 regulates AMP kinase signaling and insulin responsiveness of glucose metabolism in skeletal muscle

doi: 10.1152/ajpendo.00147.2017

Figure Lengend Snippet: Antibodies used

Article Snippet: Antibodies used are given in . table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Target Catalog No. Company PKN2 (cells) 8697 Cell Signaling Technology PAX7 27–583 Prosci DES 15200 Abcam β-Actin A5541 Sigma p-PKN1/2 (Thr 774/816 ) 2611 Cell Signaling Technology p-Akt (Ser 473 ) 9271 Cell Signaling Technology p-Akt (Thr 308 ) 4056 Cell Signaling Technology Akt 9272 Cell Signaling Technology Phospho-GSK-3α/β (Ser 21/9 ) 9331 Cell Signaling Technology GSK-3β 9315 Cell Signaling Technology p-TBC1D4 8619 Cell Signaling Technology TBC1D4 07–741 EMB Millipore p-mTOR (Ser 2448 ) 5536 Cell Signaling Technology mTOR (7C10) 2983 Cell Signaling Technology p-AMPK (Thr 172 ) 2531 Cell Signaling Technology AMPK 2532 Cell Signaling Technology p-ACC (Ser 79 ) 3661 Cell Signaling ACC 3676 Cell Signaling Technology Fyn sc-16 Santa Cruz Biotechnology GAPDH 25778 Santa Cruz Biotechnology p-S6 (Ser 235/236 ) 2211 Cell Signaling Technology S6 2317 Cell Signaling Technology PKN2 (mouse muscle) 2612 Cell Signaling Technology Open in a separate window PKN2, protein kinase N2; DES, desmin; mTOR, mammalian target of rapamycin; AMPK, AMP-activated protein kinase; ACC, acetyl-CoA carboxylase; p, phosphorylated.

Techniques:

Journal: American Journal of Physiology - Endocrinology and Metabolism

Article Title: Protein kinase N2 regulates AMP kinase signaling and insulin responsiveness of glucose metabolism in skeletal muscle

doi: 10.1152/ajpendo.00147.2017

Figure Lengend Snippet: Protein kinase N2 (PKN2) knockdown decreases insulin responsiveness of glucose metabolism in skeletal muscle. A and B: mRNA levels of PKN2, PAX7, MYOG (myogenin), and DES (desmin) (A) and protein abundance of PKN2, PAX7, and DES (B) in small-interfering (si)RNA-treated primary HSMCs. C: representative bright-field images of siRNA-treated primary human skeletal muscle cells (HSMC). Scale bar, 100 µm. D–F: basal and insulin-stimulated (120 nM) glucose uptake (D), incorporation into glycogen (E), and oxidation (F) in siRNA-treated primary HSMC. Open bars, scrambled (SCR); black bars, small-interfering (si)PKN2. *PKN2 effect, P < 0.05; #insulin main effect, P < 0.05. Results are means ± SE for n = 5 biological replicates.

Article Snippet: Antibodies used are given in . table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Target Catalog No. Company PKN2 (cells) 8697 Cell Signaling Technology PAX7 27–583 Prosci DES 15200 Abcam β-Actin A5541 Sigma p-PKN1/2 (Thr 774/816 ) 2611 Cell Signaling Technology p-Akt (Ser 473 ) 9271 Cell Signaling Technology p-Akt (Thr 308 ) 4056 Cell Signaling Technology Akt 9272 Cell Signaling Technology Phospho-GSK-3α/β (Ser 21/9 ) 9331 Cell Signaling Technology GSK-3β 9315 Cell Signaling Technology p-TBC1D4 8619 Cell Signaling Technology TBC1D4 07–741 EMB Millipore p-mTOR (Ser 2448 ) 5536 Cell Signaling Technology mTOR (7C10) 2983 Cell Signaling Technology p-AMPK (Thr 172 ) 2531 Cell Signaling Technology AMPK 2532 Cell Signaling Technology p-ACC (Ser 79 ) 3661 Cell Signaling ACC 3676 Cell Signaling Technology Fyn sc-16 Santa Cruz Biotechnology GAPDH 25778 Santa Cruz Biotechnology p-S6 (Ser 235/236 ) 2211 Cell Signaling Technology S6 2317 Cell Signaling Technology PKN2 (mouse muscle) 2612 Cell Signaling Technology Open in a separate window PKN2, protein kinase N2; DES, desmin; mTOR, mammalian target of rapamycin; AMPK, AMP-activated protein kinase; ACC, acetyl-CoA carboxylase; p, phosphorylated.

Techniques:

Journal: American Journal of Physiology - Endocrinology and Metabolism

Article Title: Protein kinase N2 regulates AMP kinase signaling and insulin responsiveness of glucose metabolism in skeletal muscle

doi: 10.1152/ajpendo.00147.2017

Figure Lengend Snippet: Antibodies used

Article Snippet: PKN2 (cells) , 8697 , Cell Signaling Technology.

Techniques:

Journal: American Journal of Physiology - Endocrinology and Metabolism

Article Title: Protein kinase N2 regulates AMP kinase signaling and insulin responsiveness of glucose metabolism in skeletal muscle

doi: 10.1152/ajpendo.00147.2017

Figure Lengend Snippet: Primers used

Article Snippet: PKN2 (cells) , 8697 , Cell Signaling Technology.

Techniques:

Journal: American Journal of Physiology - Endocrinology and Metabolism

Article Title: Protein kinase N2 regulates AMP kinase signaling and insulin responsiveness of glucose metabolism in skeletal muscle

doi: 10.1152/ajpendo.00147.2017

Figure Lengend Snippet: Protein kinase N2 (PKN2) knockdown decreases insulin responsiveness of glucose metabolism in skeletal muscle. A and B: mRNA levels of PKN2, PAX7, MYOG (myogenin), and DES (desmin) (A) and protein abundance of PKN2, PAX7, and DES (B) in small-interfering (si)RNA-treated primary HSMCs. C: representative bright-field images of siRNA-treated primary human skeletal muscle cells (HSMC). Scale bar, 100 µm. D–F: basal and insulin-stimulated (120 nM) glucose uptake (D), incorporation into glycogen (E), and oxidation (F) in siRNA-treated primary HSMC. Open bars, scrambled (SCR); black bars, small-interfering (si)PKN2. *PKN2 effect, P < 0.05; #insulin main effect, P < 0.05. Results are means ± SE for n = 5 biological replicates.

Article Snippet: PKN2 (cells) , 8697 , Cell Signaling Technology.

Techniques: Knockdown, Quantitative Proteomics

Journal: American Journal of Physiology - Endocrinology and Metabolism

Article Title: Protein kinase N2 regulates AMP kinase signaling and insulin responsiveness of glucose metabolism in skeletal muscle

doi: 10.1152/ajpendo.00147.2017

Figure Lengend Snippet: PKN2 knockdown increases TBC1D4 phosphorylation in HSMCs. A:Western blot analysis of PKN2, Akt, GSK-3, and TBC1D4 protein and phosphorylation from basal and insulin-stimulated (120 nM; 15 min) primary HSMCs (representative immunoblot from n = 5 biological replicates). B: Western blot analysis of PKN2 and Akt protein and phosphorylation in mouse quadriceps muscle 15 min following saline or insulin (5 IU/kg ip) injection (representative immunoblot from n = 4 mice). C: quantification of phosphorylated (p)-TBC1D4 Ser318 abundance in basal and insulin-stimulated primary HSMCs. Open bars, SCR; black bars, siPKN2. *PKN2 effect, P < 0.05; #insulin main effect, P < 0.05. Results are means ± SE for n = 5 biological replicates.

Article Snippet: PKN2 (cells) , 8697 , Cell Signaling Technology.

Techniques: Knockdown, Phospho-proteomics, Western Blot, Saline, Injection

Journal: American Journal of Physiology - Endocrinology and Metabolism

Article Title: Protein kinase N2 regulates AMP kinase signaling and insulin responsiveness of glucose metabolism in skeletal muscle

doi: 10.1152/ajpendo.00147.2017

Figure Lengend Snippet: PKN2 knockdown increases AMP-activated protein kinase (AMPK) signaling. A: Western blot analysis of p-AMPK Thr172, AMPK, and p-ACC Ser79 in primary HSMCs incubated in the absence or presence of insulin (120 nM; 15 min) (representative immunoblot from n = 5 biological replicates). B and C: quantification of p-AMPK Thr172 (B) and p-ACC Ser79 abundance (C). D: Western blot analysis of p-ACC Ser79 abundance in PKN2 siRNA-treated human embryonic kidney-293 cells overexpressing constitutively active Fyn kinase (caFyn; representative immunoblot from n = 3 biological replicates). Open bars, SCR; black bars, siPKN2. *PKN2 post hoc effect. Results are means ± SE for n = 5 biological replicates.

Article Snippet: PKN2 (cells) , 8697 , Cell Signaling Technology.

Techniques: Knockdown, Western Blot, Incubation

Journal: American Journal of Physiology - Endocrinology and Metabolism

Article Title: Protein kinase N2 regulates AMP kinase signaling and insulin responsiveness of glucose metabolism in skeletal muscle

doi: 10.1152/ajpendo.00147.2017

Figure Lengend Snippet: PKN2 knockdown increases fatty acid oxidation and incorporation into triglycerides (TAG). Palmitate oxidation (A) and incorporation (B) into lipid species in siRNA-treated primary HSMC incubated in the absence or presence of without 5-aminoimidazole-4-carboxamide-1-β-4-ribofuranoside (AICAR; 2 mM). mRNA level of peroxisome proliferator-activated receptor (PPAR)-γ coactivator-1α (PGC-1α; C) and sterol regulatory element-binding protein-1c (SREBP-1c; D) target genes in siRNA-treated primary HSMCs. Open bars, SCR; black bars, siPKN2. *PKN2 post hoc effect, P < 0.05; #AICAR main effect, P < 0.05. Results are means ± SE for n = 5 biological replicates. CPT-1β, carnitine palmitoyltransferase-1β;FABP3, fatty acid-binding protein 3; PDK4, pyruvate dehydrogenase kinase 4; SCD1, stearoyl-CoA desaturase-1; ACC2, acetyl-CoA carboxylase 2; DGAT1, diacylglycerol O-acyltransferase 1; GPAT1, glycerol-3-phosphate acyltransferase 1.

Article Snippet: PKN2 (cells) , 8697 , Cell Signaling Technology.

Techniques: Knockdown, Incubation, Binding Assay

Journal: American Journal of Physiology - Endocrinology and Metabolism

Article Title: Protein kinase N2 regulates AMP kinase signaling and insulin responsiveness of glucose metabolism in skeletal muscle

doi: 10.1152/ajpendo.00147.2017

Figure Lengend Snippet: PKN2 knockdown decreases mammalian target of rapamycin (mTOR) signaling and protein synthesis. A:Western blot analysis of p-mTOR Ser2448, mTOR, p-S6 Ser235/236, and S6 in primary HSMCs incubated in the absence or presence of insulin (120 nM; 15 min) (representative immunoblot from n = 5 biological replicates). B and C: quantification of p-mTOR Ser2448 (B) and p-S6 Ser235/236 abundance (C). D: protein synthesis in siRNA-treated primary HSMCs. Open bars, SCR; black bars, siPKN2. *PKN2 effect, P < 0.05; #insulin main effect, P < 0.05. Results are means ± SE for n = 5 biological replicates.

Article Snippet: PKN2 (cells) , 8697 , Cell Signaling Technology.

Techniques: Knockdown, Western Blot, Incubation

Journal: American Journal of Physiology - Endocrinology and Metabolism

Article Title: Protein kinase N2 regulates AMP kinase signaling and insulin responsiveness of glucose metabolism in skeletal muscle

doi: 10.1152/ajpendo.00147.2017

Figure Lengend Snippet: PKN2 silencing in vivo decreases glucose uptake and activates AMPK. Contralateral tibialis anterior muscles were electroporated with shRNA targeting PKN2 or scrambled control. Seven days following electroporation, 4-h fasted mice were administered an oral glucose load (3 g/kg), followed by ip injection of [3H]deoxyglucose. Tibialis anterior muscle was harvested 2 h following the oral glucose challenge and analyzed for PKN2, P-AMPKThr172, AMPK, P-ACC Ser79, and ACC protein abundance (representative immunoblot from n = 7 mice) (A), quantification of PKN2 protein abundance (B), in vivo glucose uptake (C), intramuscular glycogen content (D), quantification of p-AMPK Thr172 abundance (E), and p-ACC Ser79 abundance (F) in PKN2 shRNA-treated mouse tibialis anterior muscle. *Paired t-test, P < 0.05. Results are means ± SE for n = 7 mice. AU, arbitrary units.

Article Snippet: PKN2 (cells) , 8697 , Cell Signaling Technology.

Techniques: In Vivo, Muscles, shRNA, Control, Electroporation, Injection, Quantitative Proteomics, Western Blot