Western Blot:Article Title: Osteoblast-derived osteomodulin restrains osteoclastogenesis via ITGB8/RRM2-mediated reduction of mitochondrial respiration and mitochondrial ATP production
Article Snippet: Protein bands were visualized using the Amersham Imager 600 (GE) and quantified using ImageJ. .. Western blotting (WB) was performed with the following antibodies: anti-GAPDH (1:10,000, 60004-1-IG; Proteintech), anti-β-actin (1:10,000, 66009-1-IG; Proteintech), anti-cathepsin K (CTSK) (1:100, sc-48353; Santa Cruz), anti-NFATC1 (1:100, sc-7294; Santa Cruz), anti-matrix metallopeptidase 9 (MMP9) (1:1,000, 10375-2-AP; Proteintech), anti-RRM2 (1:100, sc-81850; Santa Cruz), anti-ITGB8 (1:1,000, 29775-1-AP; Proteintech), anti-OMD (1:1,000, ab154249; Abcam), anti-RUNX2 (1:1,000, 20700-1-AP; Proteintech), anti-collagen type I (1:1,000, 14695-1-AP; Proteintech), anti-ATP Synthase F1 Subunit Alpha (ATP5A1) (1:10,000, 14676-1-AP; Proteintech), anti-ubiquinol-cytochrome c reductase core protein 2 (UQCRC2) (1:10,000, 67547-1-Ig; Proteintech), anti-succinate dehydrogenase subunits B (SDHB) (1:10,000, 10620-1-AP; Proteintech), anti-cytochrome oxidase subunit IV (MTCO4) (1:1,000, 11242-1-AP; Proteintech), anti-NADH-ubiquinone oxidoreductase ASHI subunit (NDUFB8) (1:10,000, 14794-1-AP; Proteintech), anti-FLAG (1:10,000, 20543-1-AP; Proteintech), anti-HA (1:10,000, 51064-2-AP; Proteintech), anti-YAP (1:1,000, ET1608-30; HUABIO), anti-p-YAP (S127) (1:1,000, ET1611–69; HUABIO), anti-Smad2 (1:1,000, ET1604-22; HUABIO), anti-p-Smad2 (1:5,000, ET1612-32; HUABIO), anti-Smad3 (1:5,000, ET1607-41; HUABIO), anti-p-Smad3 (1:2,000, ET1609-41; HUABIO), anti-AKT (1:1,000, #9272, Cell Signaling Technology), anti-p-AKT (1:2,000, #4060, Cell Signaling Technology), anti-ERK (1:1,000, #9102, Cell Signaling Technology), anti-p-ERK (1:1,000, #4695, Cell Signaling Technology), anti-p38 (1:1,000, #9212, Cell Signaling Technology), anti-p-p38 (1:1,000, #4511, Cell Signaling Technology), anti-JNK (1:5,000, 51153-1-AP; Proteintech) and anti-p-JNK (1:1,000, 80024-1-RR; Proteintech). ..
Article Title: MCM8 interacts with DDX5 to promote R-loop resolution
Article Snippet: .. The primary antibodies used for immunoprecipitation (IP) and western blotting (WB) were rabbit anti-MCM8 (5 μg for IP, 1:1000 dilution for WB; Proteintech 16451-1-AP), rabbit anti-DDX5 (5 μg for IP, 1:1000 dilution for WB; Abcam ab21696), rabbit anti-DHX9 (5 μg for IP, 1:900 dilution for WB; Abcam ab26271), rabbit anti-HA (5 μg for IP, 1:1000 dilution for WB; Cell Signaling Technology 3724), mouse anti-MYC (1:1000 dilution for WB; Cell Signaling Technology 2276), mouse anti-FLAG (1:1000 dilution for WB; Sigma-Aldrich F1804), mouse anti-S9.6 (5 μg for IP; Kerafast ENH001), normal rabbit IgG (5 μg for IP; Cell Signaling Technology 3700), normal mouse IgG (5 μg for IP; Proteintech B900620), and mouse anti-β-actin (1:5000 dilution for WB; Proteintech 66009-1-Ig). .. The primary antibodies used for immunostaining were goat anti-STELLA (1:50 dilution; Novus Biologicals AF2566), goat anti-DDX4 (1:50 dilution; Novus Biologicals AF2030), rabbit anti-FOXL2 (1:250 dilution; Abcam ab246511), rabbit anti-SOX9 (1:200 dilution; Millipore AB5535), rabbit anti-cleaved PARP1 (1:100 dilution; Cell Signaling Technology 94885), rabbit anti-Ki67 (1:200 dilution; Abcam ab15580), rabbit anti-Cyclin B1 (1:200 dilution; Cell Signaling Technology 4138), rabbit anti-53BP1 (1:500 dilution; Novus Biologicals NB100-304), rabbit anti-γH2AX (1:50 dilution; Cell Signaling Technology 9718), rabbit anti-pATM (1:100 dilution; Cell Signaling Technology 5883), rabbit anti-pSer2 Pol II (1:100 dilution; Abcam Ab193468), rabbit anti-pSer5 Pol II (1:100 dilution; Abcam Ab193467), rabbit anti-5mC (1:100 dilution; Abcam Ab214727), rabbit anti-H3K9me2 (1:100 dilution; Millipore 07-441), rabbit anti-H3K27me3 (1:100 dilution; Millipore 07-449), rabbit anti-S9.6 (1:200 dilution; Kerafast Kf-Ab01137-23.0), goat anti-S9.6 (1:400 dilution; Kerafast Kf-Ab01137-24.1), mouse anti-Fibrillarin (1:500 dilution; Abcam ab4566), rabbit anti-DDX5 (1:400 dilution; Abcam ab21696), rabbit anti-DHX9 (1:400 dilution; Abcam ab26271), and mouse anti-HA (1:200 dilution; Cell Signaling Technology 2367).
Article Title: Reversible ubiquitination regulates HSF1 phase separation in response to proteotoxic stress.
Article Snippet: This is a PDF of an article that has undergone enhancements after acceptance, such as the addition of a cover page and metadata, and formatting for readability.. This version will undergo additional copyediting, typesetting and review before it is published in its final form.. As such, this version is no longer the Accepted Manuscript, but it is not yet the definitive Version of Record; we are providing this early version to give early visibility of the article.
Article Title: Generation and characterization of a knock-in mouse model for Spastic Tetraplegia, Thin Corpus Callosum, and Progressive Microcephaly (SPATCCM)
Article Snippet: Proteins were electrophoresed through 8% SDS-polyacrylamide gels and transferred to Immobilon P membrane (Millipore). .. Western blotting (WB) was performed following standard protocols using rabbit anti-SLC1A4 (Proteintech Cat# 13067-2-AP, RRID:AB_2190604 or Cell Signaling Technology Cat# 8442, RRID:AB_10828382) and mouse anti-beta-tubulin-III (3F3-G2) (Santa Cruz Biotechnology Cat# sc-53140, RRID:AB_793543). .. Following ECL (BioRad Clarity ECL substrate), blots were imaged using an Azure 300 imager and quanti ed using AzureSpot Pro (Azure Biosystems).
Article Title: Osteoblast-derived osteomodulin restrains osteoclastogenesis via ITGB8/RRM2-mediated reduction of mitochondrial respiration and mitochondrial ATP production.
Article Snippet: Protein bands were visualized using the Amersham Imager 600 (GE) and quantified using ImageJ. .. Western blotting (WB) was performed with the following antibodies: anti-GAPDH (1:10,000, 60004-1-IG; Proteintech), anti-β-actin (1:10,000, 66009-1-IG; Proteintech), anti-cathepsin K (CTSK) (1:100, sc-48353; Santa Cruz), anti-NFATC1 (1:100, sc-7294; Santa Cruz), anti-matrix metallopeptidase 9 (MMP9) (1:1,000, 10375-2-AP; Proteintech), anti-RRM2 (1:100, sc-81850; Santa Cruz), anti-ITGB8 (1:1,000, 29775-1-AP; Proteintech), anti-OMD (1:1,000, ab154249; Abcam), antiRUNX2 (1:1,000, 20700-1-AP; Proteintech), anti-collagen type I (1:1,000, 14695-1-AP; Proteintech), anti-ATP Synthase F1 Subunit Alpha (ATP5A1) (1:10,000, 14676-1-AP; Proteintech), anti-ubiquinol-cytochrome c reductase core protein 2 (UQCRC2) (1:10,000, 67547-1-Ig; Proteintech), anti-succinate dehydrogenase subunits B (SDHB) (1:10,000, 10620-1-AP; Proteintech), anti-cytochrome oxidase subunit IV (MTCO4) (1:1,000, 11242-1-AP; Proteintech), anti-NADH-ubiquinone oxidoreductase ASHI subunit (NDUFB8) (1:10,000, 14794-1-AP; Proteintech), anti-FLAG (1:10,000, 20543- 1-AP; Proteintech), anti-HA (1:10,000, 51064-2-AP; Proteintech), anti-YAP (1:1,000, ET1608-30; HUABIO), anti-p-YAP (S127) (1:1,000, ET1611–69; HUABIO), anti-Smad2 (1:1,000, ET1604-22; HUABIO), anti-p-Smad2 (1:5,000, ET1612-32; HUABIO), anti-Smad3 (1:5,000, ET1607-41; HUABIO), anti-p-Smad3 (1:2,000, ET1609-41; HUABIO), anti-AKT (1:1,000, #9272, Cell Signaling Technology), anti-p-AKT (1:2,000, #4060, Cell Signaling Technology), anti-ERK (1:1,000, #9102, Cell Signaling Technology), anti-p-ERK (1:1,000, #4695, Cell Signaling Technology), anti-p38 (1:1,000, #9212, Cell Signaling Technology), anti-p-p38 (1:1,000, #4511, Cell Signaling Technology), anti-JNK (1:5,000, 51153-1-AP; Proteintech) and anti-p-JNK (1:1,000, 80024-1-RR; Proteintech). ..
Article Title: Bmal1 upregulates ATG5 expression to promote autophagy in skin cutaneous melanoma.
Article Snippet: .. Western blotting (WB) was performed based on the previously described protocol [22], using the following antibodies: anti-ARNTL (Proteintech, Cat#14268–1-AP), anti-P62 (Cat#: 29503–1-AP), antiLC3 (Cat#:14600–1-AP), anti-ATG5 (HUABIO, Cat#: ET1611–38), and anti-β-actin (Bioworld, Cat#: AP0060). .. Secondary anti-rabbit and antimouse antibodies were obtained from Bioworld Technology.
Immunoprecipitation:Article Title: MCM8 interacts with DDX5 to promote R-loop resolution
Article Snippet: .. The primary antibodies used for immunoprecipitation (IP) and western blotting (WB) were rabbit anti-MCM8 (5 μg for IP, 1:1000 dilution for WB; Proteintech 16451-1-AP), rabbit anti-DDX5 (5 μg for IP, 1:1000 dilution for WB; Abcam ab21696), rabbit anti-DHX9 (5 μg for IP, 1:900 dilution for WB; Abcam ab26271), rabbit anti-HA (5 μg for IP, 1:1000 dilution for WB; Cell Signaling Technology 3724), mouse anti-MYC (1:1000 dilution for WB; Cell Signaling Technology 2276), mouse anti-FLAG (1:1000 dilution for WB; Sigma-Aldrich F1804), mouse anti-S9.6 (5 μg for IP; Kerafast ENH001), normal rabbit IgG (5 μg for IP; Cell Signaling Technology 3700), normal mouse IgG (5 μg for IP; Proteintech B900620), and mouse anti-β-actin (1:5000 dilution for WB; Proteintech 66009-1-Ig). .. The primary antibodies used for immunostaining were goat anti-STELLA (1:50 dilution; Novus Biologicals AF2566), goat anti-DDX4 (1:50 dilution; Novus Biologicals AF2030), rabbit anti-FOXL2 (1:250 dilution; Abcam ab246511), rabbit anti-SOX9 (1:200 dilution; Millipore AB5535), rabbit anti-cleaved PARP1 (1:100 dilution; Cell Signaling Technology 94885), rabbit anti-Ki67 (1:200 dilution; Abcam ab15580), rabbit anti-Cyclin B1 (1:200 dilution; Cell Signaling Technology 4138), rabbit anti-53BP1 (1:500 dilution; Novus Biologicals NB100-304), rabbit anti-γH2AX (1:50 dilution; Cell Signaling Technology 9718), rabbit anti-pATM (1:100 dilution; Cell Signaling Technology 5883), rabbit anti-pSer2 Pol II (1:100 dilution; Abcam Ab193468), rabbit anti-pSer5 Pol II (1:100 dilution; Abcam Ab193467), rabbit anti-5mC (1:100 dilution; Abcam Ab214727), rabbit anti-H3K9me2 (1:100 dilution; Millipore 07-441), rabbit anti-H3K27me3 (1:100 dilution; Millipore 07-449), rabbit anti-S9.6 (1:200 dilution; Kerafast Kf-Ab01137-23.0), goat anti-S9.6 (1:400 dilution; Kerafast Kf-Ab01137-24.1), mouse anti-Fibrillarin (1:500 dilution; Abcam ab4566), rabbit anti-DDX5 (1:400 dilution; Abcam ab21696), rabbit anti-DHX9 (1:400 dilution; Abcam ab26271), and mouse anti-HA (1:200 dilution; Cell Signaling Technology 2367).
Immunofluorescence:Article Title: Reversible ubiquitination regulates HSF1 phase separation in response to proteotoxic stress.
Article Snippet: This is a PDF of an article that has undergone enhancements after acceptance, such as the addition of a cover page and metadata, and formatting for readability.. This version will undergo additional copyediting, typesetting and review before it is published in its final form.. As such, this version is no longer the Accepted Manuscript, but it is not yet the definitive Version of Record; we are providing this early version to give early visibility of the article.
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