primary mouse polyclonal antibody against β actin  (Santa Cruz Biotechnology)

Journal: Biochemical and biophysical research communications

Article Title: The stiffness and collagen control differentiation of osteoclasts with an altered expression of c-Src in podosome.

doi: 10.1016/j.bbrc.2024.149636

Figure Lengend Snippet: Fig. 1. RANKL induced osteoclast differentiation and mRNA expression of osteoclast markers in Raw264.7 cells. Raw264.7 cells were treated with or without 100 ng/mL of sRANKL for 4 days. (A) Osteoclasts were stained with TRAP. The scale bar indicates 200 μm. (B) Nuclei and actin were stained with DAPI (blue) and phalloidin (red), respectively, on the indicated days. The scale bar indicates 50 μm. (C and D) Total RNA was extracted, and the mRNA expression of the osteoclast marker genes Nfatc1 (C) and Ctsk (D) was observed. β-Actin was used as a normalized gene. Data are expressed as the mean ± SEM from three independent ex periments. Asterisks indicate a significant difference between the groups: **P < 0.01 and ***P < 0.001. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Primary antibodies against mouse β-actin (Santa Cruz Biotechnology Inc., Dallas, TX, USA) and Src (Cell Signaling Technology Inc., Danvers, MA, USA) were used.

Techniques: Expressing, Staining, Marker

Journal: Biochemical and biophysical research communications

Article Title: The stiffness and collagen control differentiation of osteoclasts with an altered expression of c-Src in podosome.

doi: 10.1016/j.bbrc.2024.149636

Figure Lengend Snippet: Fig. 2. Podosome components were upregulated during RANKL-induced osteoclast differentiation in Raw264.7 cells. (A–C) Total RNA was extracted, and the mRNA expression of the podosome-related factors Itgαv (A), Itgβ3 (B), and Vcl (C) were analyzed by RT-qPCR. β-actin was used as a normalized gene. The data are expressed as the mean ± SEM from a representative of three independent experiments. Asterisks indicate a significant difference between the groups: *P < 0.05, **P < 0.01, and ***P < 0.001. (D) Total protein was extracted and subjected to Western blotting to detect Src and β-actin proteins. β-Actin was used as a reference. (E) The relative intensity of Src shown in (D) was measured.

Article Snippet: Primary antibodies against mouse β-actin (Santa Cruz Biotechnology Inc., Dallas, TX, USA) and Src (Cell Signaling Technology Inc., Danvers, MA, USA) were used.

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Journal: Biochemical and biophysical research communications

Article Title: The stiffness and collagen control differentiation of osteoclasts with an altered expression of c-Src in podosome.

doi: 10.1016/j.bbrc.2024.149636

Figure Lengend Snippet: Fig. 3. RGD sequences promote osteoclast differentiation with enhanced expression of podosome-related factors. Raw264.7 cells were treated with 100 ng/mL of sRANKL for 4 days on non-coated (RGD-), BSA-coated (RGD-), or collagen-coated (RGD+) plates. (A) Osteoclasts were stained for TRAP on the indicated days. The scale bar indicates 200 μm. (B–F) Total RNA was extracted, and the mRNA expression of the osteoclast marker genes Nfatc1 (B) and Ctsk (C) and the podosome- related factors Itgαv (D), Itgβ3 (E), and Vcl (F) were analyzed by RT-qPCR. β-Actin was used as a normalized gene. Data are expressed as mean ± SEM from three independent experiments. Asterisks indicate significant differences between the groups: *P < 0.05, **P < 0.01, and ***P < 0.001. (D) Total protein was extracted and subjected to Western blotting to detect Src and β-actin. β-Actin was used as a reference. (E) The relative intensity of Src, shown in (D), was measured.

Article Snippet: Primary antibodies against mouse β-actin (Santa Cruz Biotechnology Inc., Dallas, TX, USA) and Src (Cell Signaling Technology Inc., Danvers, MA, USA) were used.

Techniques: Expressing, Staining, Marker, Quantitative RT-PCR, Western Blot

Journal: Biochemical and biophysical research communications

Article Title: The stiffness and collagen control differentiation of osteoclasts with an altered expression of c-Src in podosome.

doi: 10.1016/j.bbrc.2024.149636

Figure Lengend Snippet: Fig. 4. Extracellular matrix stiffness regulates osteoclast differentiation and expression of podosome-related factors. Raw264.7 cells were treated with 100 ng/mL of sRANKL for 4 days on soft collagen gel or hard collagen-coated plates. (A) Osteoclasts were stained for TRAP on the indicated days. The scale bar indicates 200 μm. (B–F) Total RNA was extracted, and the mRNA expression of the osteoclast marker genes Nfatc1 (B) and Ctsk (C) and the podosome-related factors Itgαv (D), Itgβ3 (E), and Vcl (F) were analyzed by RT-qPCR. β-Actin was used as a normalized gene. Data are expressed as the mean ± SEM from three independent experiments. Asterisks indicate significant differences between the two groups: *P < 0.05, **P < 0.01, and ***P < 0.001. (D) Total protein was extracted and subjected to Western blotting to detect Src and β-actin. β-Actin was used as a reference. (E) The relative intensity of Src, shown in (D), was measured.

Article Snippet: Primary antibodies against mouse β-actin (Santa Cruz Biotechnology Inc., Dallas, TX, USA) and Src (Cell Signaling Technology Inc., Danvers, MA, USA) were used.

Techniques: Expressing, Staining, Marker, Quantitative RT-PCR, Western Blot