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Small interfering RNA (siRNA)-mediated knockdown of BET1L was efficacious in WT rats. (A) A representative Western blot image showing the relative levels of Bet1L protein in using muscle homogenates. Bet1L siRNA was prepared at the final concentration of 5 mM and injected into the hind limb (tibialis anterior) muscles. Three control groups were prepared: scrambled siRNA, vehicle only <t>(Oligofectamine),</t> and no-injection sham. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as an internal control. In this electrophoresis, two muscle samples from different rats were loaded for each treatment group. (B) The densitometric analysis of proteins revealed that Bet1L siRNA injection significantly reduced its protein level up to ~18% when compared to the level in the non-injection control rats. * P < 0.05 vs. three control groups, n = 6 each. (C) Representative images of immunohistochemistry showing the decreased Bet1L protein at the neuromuscular junctions (NMJs) in the Bet1L siRNA-injected muscle. AChR, acetylcholine receptor. Scale bar = 10 mm. (D) The number of Bet1L-positive NMJs was significantly decreased after 1 week of Bet1L siRNA injection in WT rats. ** P < 0.01 vs. three control groups, n = 4 each.
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Small interfering RNA (siRNA)-mediated knockdown of BET1L was efficacious in WT rats. (A) A representative Western blot image showing the relative levels of Bet1L protein in using muscle homogenates. Bet1L siRNA was prepared at the final concentration of 5 mM and injected into the hind limb (tibialis anterior) muscles. Three control groups were prepared: scrambled siRNA, vehicle only <t>(Oligofectamine),</t> and no-injection sham. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as an internal control. In this electrophoresis, two muscle samples from different rats were loaded for each treatment group. (B) The densitometric analysis of proteins revealed that Bet1L siRNA injection significantly reduced its protein level up to ~18% when compared to the level in the non-injection control rats. * P < 0.05 vs. three control groups, n = 6 each. (C) Representative images of immunohistochemistry showing the decreased Bet1L protein at the neuromuscular junctions (NMJs) in the Bet1L siRNA-injected muscle. AChR, acetylcholine receptor. Scale bar = 10 mm. (D) The number of Bet1L-positive NMJs was significantly decreased after 1 week of Bet1L siRNA injection in WT rats. ** P < 0.01 vs. three control groups, n = 4 each.
Oligofectamine™ Transfection Reagent, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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oligofectamine™ transfection reagent - by Bioz Stars, 2026-10
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Small interfering RNA (siRNA)-mediated knockdown of BET1L was efficacious in WT rats. (A) A representative Western blot image showing the relative levels of Bet1L protein in using muscle homogenates. Bet1L siRNA was prepared at the final concentration of 5 mM and injected into the hind limb (tibialis anterior) muscles. Three control groups were prepared: scrambled siRNA, vehicle only (Oligofectamine), and no-injection sham. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as an internal control. In this electrophoresis, two muscle samples from different rats were loaded for each treatment group. (B) The densitometric analysis of proteins revealed that Bet1L siRNA injection significantly reduced its protein level up to ~18% when compared to the level in the non-injection control rats. * P < 0.05 vs. three control groups, n = 6 each. (C) Representative images of immunohistochemistry showing the decreased Bet1L protein at the neuromuscular junctions (NMJs) in the Bet1L siRNA-injected muscle. AChR, acetylcholine receptor. Scale bar = 10 mm. (D) The number of Bet1L-positive NMJs was significantly decreased after 1 week of Bet1L siRNA injection in WT rats. ** P < 0.01 vs. three control groups, n = 4 each.

Journal: Frontiers in Neuroscience

Article Title: Muscle-specific Bet1L knockdown induces neuromuscular denervation, motor neuron degeneration, and motor dysfunction in a rat model of familial ALS

doi: 10.3389/fnins.2025.1527181

Figure Lengend Snippet: Small interfering RNA (siRNA)-mediated knockdown of BET1L was efficacious in WT rats. (A) A representative Western blot image showing the relative levels of Bet1L protein in using muscle homogenates. Bet1L siRNA was prepared at the final concentration of 5 mM and injected into the hind limb (tibialis anterior) muscles. Three control groups were prepared: scrambled siRNA, vehicle only (Oligofectamine), and no-injection sham. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as an internal control. In this electrophoresis, two muscle samples from different rats were loaded for each treatment group. (B) The densitometric analysis of proteins revealed that Bet1L siRNA injection significantly reduced its protein level up to ~18% when compared to the level in the non-injection control rats. * P < 0.05 vs. three control groups, n = 6 each. (C) Representative images of immunohistochemistry showing the decreased Bet1L protein at the neuromuscular junctions (NMJs) in the Bet1L siRNA-injected muscle. AChR, acetylcholine receptor. Scale bar = 10 mm. (D) The number of Bet1L-positive NMJs was significantly decreased after 1 week of Bet1L siRNA injection in WT rats. ** P < 0.01 vs. three control groups, n = 4 each.

Article Snippet: Scrambled sequence siRNA (Dharmacon, D-001810-10-05) was also purchased as a control. siRNA was mixed with Oligofectamine TM transfection reagent (12252-011, Thermo Fisher Scientific, Waltham, MA) and RNAase (Qiagen, Hilden, Germany) in a buffer (diluted from 5 × siRNA buffer, NC1338268, Thermo Fisher Scientific) for a final concentration of 5 μM siRNA.

Techniques: Small Interfering RNA, Knockdown, Western Blot, Concentration Assay, Injection, Muscles, Control, Electrophoresis, Immunohistochemistry