erbb4 sirna  (Santa Cruz Biotechnology)

Journal: Acta Biochimica et Biophysica Sinica

Article Title: Neuregulin-1/ErbB4 upregulates acetylcholine receptors via Akt/mTOR/p70S6K: a study in a rat model of obstetric brachial plexus palsy and in vitro

doi: 10.3724/abbs.2022158

Figure Lengend Snippet: Table 1 Sequence of primers used in real-time PCR

Article Snippet: L6 myoblasts were transfected with ErbB4 siRNA to inhibit the endogenous ErbB4 level using Lipo2000 (Invitrogen) in accordance with the manufacturer’s protocol, and NC siRNA transfection was used as a control.

Techniques: Sequencing, Real-time Polymerase Chain Reaction

Journal: Acta Biochimica et Biophysica Sinica

Article Title: Neuregulin-1/ErbB4 upregulates acetylcholine receptors via Akt/mTOR/p70S6K: a study in a rat model of obstetric brachial plexus palsy and in vitro

doi: 10.3724/abbs.2022158

Figure Lengend Snippet: The expression levels of NRG-1 and ErbB4 are lower in denervated IMF than in denervated biceps of the OBPP rat model The protein levels of NRG-1 and ErbB4 were more obviously decreased in denervated IMF than in denervated biceps, whereas those of ErbB2 and ErbB3 were not different between the two groups, as revealed by western blot analysis (A,B). The mRNA levels of NRG-1 and ErbB4 were reduced more significantly in denervated IMF than in denervated biceps, whereas no differences were found in the mRNA levels of ErbB2 and ErbB3 between the two denervated muscles, as revealed by real-time quantitative PCR (C). In representative confocal images (D), the density of ErbB4 protein was decreased in denervated biceps and almost disappeared in denervated IMF. Red: neurofilament; green: α-bungarotoxin; blue: ErbB4. Scale bar: 20 μm. n=6. Denervated IMF versus denervated biceps. * P<0.05, ** P<0.01. N.S. represents no significance.

Article Snippet: L6 myoblasts were transfected with ErbB4 siRNA to inhibit the endogenous ErbB4 level using Lipo2000 (Invitrogen) in accordance with the manufacturer’s protocol, and NC siRNA transfection was used as a control.

Techniques: Expressing, Western Blot, Real-time Polymerase Chain Reaction

Journal: Acta Biochimica et Biophysica Sinica

Article Title: Neuregulin-1/ErbB4 upregulates acetylcholine receptors via Akt/mTOR/p70S6K: a study in a rat model of obstetric brachial plexus palsy and in vitro

doi: 10.3724/abbs.2022158

Figure Lengend Snippet: The effects of NRG-1/ErbB4 on regulating the protein synthesis of AChR subunits in L6 myoblasts The protein levels of AChRα, AChRβ, and AChRδ were upregulated in L6 myoblasts treated with NRG-1, as revealed by western blot analysis (A). The protein levels of AChRα, AChRβ, and AChRδ were upregulated in the ErbB4-overexpressing group, as revealed by western blot analysis (B,D). The protein levels of AChRα, AChRβ, and AChRδ were downregulated in L6 myoblasts transfected with ErbB4 siRNA (C, E). The mRNA levels of AChRα, AChRβ, and AChRδ showed no significant changes in ErbB4-overexpressing (F) or ErbB4-knockdown (G) L6 myoblasts, as revealed by real-time quantitative PCR. The AChR cluster is upregulated in the ErbB4-overexpressing group and downregulated in the ErbB4-knockdown group in representative confocal images (H). Red: ErbB4; green: α-bungarotoxin; blue: nucleus. Scale bar: 20 μm. n=6. ErbB4 OV versus vector, and si-ErbB4 versus NC. * P<0.05, ** P<0.01. N.S. represents no significance.

Article Snippet: L6 myoblasts were transfected with ErbB4 siRNA to inhibit the endogenous ErbB4 level using Lipo2000 (Invitrogen) in accordance with the manufacturer’s protocol, and NC siRNA transfection was used as a control.

Techniques: Western Blot, Transfection, Real-time Polymerase Chain Reaction, Plasmid Preparation

Journal: Acta Biochimica et Biophysica Sinica

Article Title: Neuregulin-1/ErbB4 upregulates acetylcholine receptors via Akt/mTOR/p70S6K: a study in a rat model of obstetric brachial plexus palsy and in vitro

doi: 10.3724/abbs.2022158

Figure Lengend Snippet: The effects of Akt on regulating the protein synthesis of AChR subunits in L6 myoblasts The ErbB4-induced increases in AChRα, AChRβ, and AChRδ protein levels were blocked in L6 myoblasts treated with MK-2206 (Akt inhibitor) but were not affected by treatment with PD98059 (ERK inhibitor) or SB203580 (p38 inhibitor) (A–D). The AChF cluster is downregulated in vector and ErbB4 OV groups treated with MK-2206, but no changes are observed in either of the groups treated with PD98059 or SB203580 in representative confocal images (E). Green: ErbB4; blue: nucleus. Scale bar: 20 μm. n=6. ErbB4 OV versus Vector, ErbB4 OV+MK-2206 versus Vector+MK-2206, ErbB4 OV+PD98059 versus Vector+PD98059, and ErbB4 OV+SB203580 versus Vector+SB203580. ** P<0.01. N.S. represents no significance.

Article Snippet: L6 myoblasts were transfected with ErbB4 siRNA to inhibit the endogenous ErbB4 level using Lipo2000 (Invitrogen) in accordance with the manufacturer’s protocol, and NC siRNA transfection was used as a control.

Techniques: Plasmid Preparation

Journal: Acta Biochimica et Biophysica Sinica

Article Title: Neuregulin-1/ErbB4 upregulates acetylcholine receptors via Akt/mTOR/p70S6K: a study in a rat model of obstetric brachial plexus palsy and in vitro

doi: 10.3724/abbs.2022158

Figure Lengend Snippet: The effects of NRG-1/ErbB4 on positively regulating the phosphorylation of the Akt/mTOR/p70S6K signaling pathway The phosphorylation ratios of p-Akt/Akt, p-mTOR/mTOR, and p-p70S6K/p70S6K were increased in L6 myoblasts that overexpressed ErbB4 and decreased in L6 myoblasts transfected with ErbB4 siRNA (A–D). The effects of ErbB4 on upregulating the phosphorylation ratios of p-Akt/Akt, p-mTOR/mTOR, and p-p70S6K/p70S6K protein levels were blocked in L6 myoblasts overexpressing ErbB4 or vector with MK-2206 treatment (E–H). n=6. ErbB4 OV versus vector, si-ErbB4 versus NC, and ErbB4 OV+MK-2206 versus vector+MK-2206. ** P<0.01. N.S. represents no significance.

Article Snippet: L6 myoblasts were transfected with ErbB4 siRNA to inhibit the endogenous ErbB4 level using Lipo2000 (Invitrogen) in accordance with the manufacturer’s protocol, and NC siRNA transfection was used as a control.

Techniques: Transfection, Plasmid Preparation

Journal: Acta Biochimica et Biophysica Sinica

Article Title: Neuregulin-1/ErbB4 upregulates acetylcholine receptors via Akt/mTOR/p70S6K: a study in a rat model of obstetric brachial plexus palsy and in vitro

doi: 10.3724/abbs.2022158

Figure Lengend Snippet: The effects of NRG-1/ErbB4 on regulating the protein synthesis of AChR subunits via mTOR in L6 myoblasts The effects of ErbB4 on increasing AChRα, AChRβ, and AChRδ protein levels are blocked in L6 myoblasts treated with rapamycin (A,B). The effects of ErbB4 on increasing the phosphorylation ratios of p-mTOR/mTOR and p-p70S6K/p70S6K were blocked in L6 myoblasts treated with rapamycin (C,D). AChR clusters are decreased in L6 myoblasts overexpressing ErbB4 and vector with rapamycin treatment in representative confocal images (E). Green: α-bungarotoxin; blue: nucleus. n=6. ErbB4 OV versus vector, and ErbB4 OV+rapamycin versus vector+rapamycin. ** P<0.01. N.S. represents no significance.

Article Snippet: L6 myoblasts were transfected with ErbB4 siRNA to inhibit the endogenous ErbB4 level using Lipo2000 (Invitrogen) in accordance with the manufacturer’s protocol, and NC siRNA transfection was used as a control.

Techniques: Plasmid Preparation