primary antibodies against proliferating cell nuclear antigen (pcna)  (Thermo Fisher)

Journal: Open Medicine

Article Title: Knockdown of circ_0113656 assuages oxidized low-density lipoprotein-induced vascular smooth muscle cell injury through the miR-188-3p/IGF2 pathway

doi: 10.1515/med-2023-0687

Figure Lengend Snippet: Ox-LDL induces HVSMC injury. HVSMCs were treated with various concentrations of ox-LDL (0, 25, 50, and 100 µg/mL), and cell viability was analyzed by CCK-8 (a), cell proliferation by EdU assay (b), cell invasion by transwell assay (c), cell migration by wound-healing assay (d and e), and the protein expression of PCNA and MMP2 by Western blotting analysis (f). * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: After blocking the aspecific signals using skimmed milk (Yili, Beijing, China), polyvinylidene fluoride membranes with protein bands were incubated with the primary antibodies against proliferating cell nuclear antigen (PCNA) (#13-3900; 1:5000; Thermo Fisher, Waltham, MA, USA), matrix metalloprotein 2 (MMP2) (#436000; 1:250; Thermo Fisher), IGF2 (#MA5-17096; 1:1,000; Thermo Fisher), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (#MA1-16757; 1:2,000; Thermo Fisher).

Techniques: CCK-8 Assay, EdU Assay, Transwell Assay, Migration, Wound Healing Assay, Expressing, Western Blot

Journal: Open Medicine

Article Title: Knockdown of circ_0113656 assuages oxidized low-density lipoprotein-induced vascular smooth muscle cell injury through the miR-188-3p/IGF2 pathway

doi: 10.1515/med-2023-0687

Figure Lengend Snippet: The effects of circ_0113656 on ox-LDL-induced HVSMC injury. (a) Circ_0113656 expression was detected by qRT-PCR in the blood samples of AS patients and healthy volunteers ( N = 17 and N = 13, respectively). (b) Circ_0113656 expression was detected by qRT-PCR in HVSMCs treated with ox-LDL (0, 25, 50, and 100 µg/mL). (c and d) The circular structure of circ_0113656 was identified by using RNase R, random primers, and oligo(dT) 18 primers. HVSMCs were divided into the ox-LDL group, ox-LDL + si-NC group, and ox-LDL + si-circ_0113656 group, with untreated HVSMCs as controls, and circ_0113656 expression was analyzed by qRT-PCR (e), cell viability by CCK-8 (f), cell proliferation by EdU assay (g), cell invasion by transwell assay (h), cell migration by wound-healing assay (i), and the protein expression of PCNA and MMP2 by Western blotting analysis (j). ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: After blocking the aspecific signals using skimmed milk (Yili, Beijing, China), polyvinylidene fluoride membranes with protein bands were incubated with the primary antibodies against proliferating cell nuclear antigen (PCNA) (#13-3900; 1:5000; Thermo Fisher, Waltham, MA, USA), matrix metalloprotein 2 (MMP2) (#436000; 1:250; Thermo Fisher), IGF2 (#MA5-17096; 1:1,000; Thermo Fisher), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (#MA1-16757; 1:2,000; Thermo Fisher).

Techniques: Expressing, Quantitative RT-PCR, CCK-8 Assay, EdU Assay, Transwell Assay, Migration, Wound Healing Assay, Western Blot

Journal: Open Medicine

Article Title: Knockdown of circ_0113656 assuages oxidized low-density lipoprotein-induced vascular smooth muscle cell injury through the miR-188-3p/IGF2 pathway

doi: 10.1515/med-2023-0687

Figure Lengend Snippet: The effects of circ_0113656 knockdown and miR-188-3p depletion on ox-LDL-induced HVSMC injury. HVSMCs were divided into the ox-LDL group, ox-LDL + si-NC group, ox-LDL + si-circ_0113656 group, ox-LDL + si-circ_0113656 + anti-miR-NC group, and ox-LDL + si-circ_0113656 + anti-miR-188-3p group, with untreated HVSMCs as controls. MiR-188-3p expression was analyzed by qRT-PCR (a), cell viability by CCK-8 (b), cell proliferation by EdU assay (c), cell invasion by transwell assay (d and e), cell migration by wound-healing assay (f), and the protein expression of PCNA and MMP2 by Western blotting analysis (g). * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: After blocking the aspecific signals using skimmed milk (Yili, Beijing, China), polyvinylidene fluoride membranes with protein bands were incubated with the primary antibodies against proliferating cell nuclear antigen (PCNA) (#13-3900; 1:5000; Thermo Fisher, Waltham, MA, USA), matrix metalloprotein 2 (MMP2) (#436000; 1:250; Thermo Fisher), IGF2 (#MA5-17096; 1:1,000; Thermo Fisher), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (#MA1-16757; 1:2,000; Thermo Fisher).

Techniques: Knockdown, Expressing, Quantitative RT-PCR, CCK-8 Assay, EdU Assay, Transwell Assay, Migration, Wound Healing Assay, Western Blot

Journal: Open Medicine

Article Title: Knockdown of circ_0113656 assuages oxidized low-density lipoprotein-induced vascular smooth muscle cell injury through the miR-188-3p/IGF2 pathway

doi: 10.1515/med-2023-0687

Figure Lengend Snippet: The effects of miR-188-3p and IGF2 on ox-LDL-induced HVSMC disorders. HVSMCs were divided into the ox-LDL group, ox-LDL + miR-NC group, ox-LDL + miR-188-3p group, ox-LDL + miR-188-3p + pcDNA group, and ox-LDL + miR-188-3p + IGF2 group, with mock HVSMCs as controls. IGF2 expression was analyzed by Western blotting analysis (a), cell viability by CCK-8 (b), cell proliferation by EdU assay (c), cell invasion by transwell assay (d and e), cell migration by wound-healing assay (f), and the protein expression of PCNA and MMP2 by Western blotting analysis (g). * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: After blocking the aspecific signals using skimmed milk (Yili, Beijing, China), polyvinylidene fluoride membranes with protein bands were incubated with the primary antibodies against proliferating cell nuclear antigen (PCNA) (#13-3900; 1:5000; Thermo Fisher, Waltham, MA, USA), matrix metalloprotein 2 (MMP2) (#436000; 1:250; Thermo Fisher), IGF2 (#MA5-17096; 1:1,000; Thermo Fisher), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (#MA1-16757; 1:2,000; Thermo Fisher).

Techniques: Expressing, Western Blot, CCK-8 Assay, EdU Assay, Transwell Assay, Migration, Wound Healing Assay

Journal: Open Medicine

Article Title: Knockdown of circ_0113656 assuages oxidized low-density lipoprotein-induced vascular smooth muscle cell injury through the miR-188-3p/IGF2 pathway

doi: 10.1515/med-2023-0687

Figure Lengend Snippet: The effects of IGF2 overexpression on HVSMC phenotypes. HVSMCs were transfected with pcDNA or the overexpression plasmid of IGF2, and IGF2 expression was analyzed by Western blotting analysis (a), cell viability by CCK-8 (b), cell proliferation by EdU assay (c), cell invasion by transwell assay (d), cell migration by wound-healing assay (e), and the protein expression of PCNA and MMP2 by Western blotting analysis (f). ** P < 0.01, and *** P < 0.001.

Article Snippet: After blocking the aspecific signals using skimmed milk (Yili, Beijing, China), polyvinylidene fluoride membranes with protein bands were incubated with the primary antibodies against proliferating cell nuclear antigen (PCNA) (#13-3900; 1:5000; Thermo Fisher, Waltham, MA, USA), matrix metalloprotein 2 (MMP2) (#436000; 1:250; Thermo Fisher), IGF2 (#MA5-17096; 1:1,000; Thermo Fisher), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (#MA1-16757; 1:2,000; Thermo Fisher).

Techniques: Over Expression, Transfection, Plasmid Preparation, Expressing, Western Blot, CCK-8 Assay, EdU Assay, Transwell Assay, Migration, Wound Healing Assay

Journal: Bioengineered

Article Title: Zinc finger protein 521 attenuates osteoarthritis via the histone deacetylases 4 in the nucleus

doi: 10.1080/21655979.2022.2090203

Figure Lengend Snippet: Here, Zfp521 promoted chondrocyte proliferation and inhibited apoptosis. (a) The EdU staining for the Zfp521-siRNA and vehicle siRNA chondrocytes was performed. Scale bar: 200 μm. (b) Comparison of the TUNEL staining for the Zfp521-siRNA and vehicle siRNA chondrocytes. Scale bar: 200 μm. (c-d) Quantitative analysis of the EdU and TUNEL assays between the Zfp521-siRNA group and vehicle siRNA group. (e–f) Immunohistochemical staining for PCNA and caspase-3 in the ACLT rats at nine weeks postop. Black scale bar: 50 μm. Blue scale bar: 15 μm. (g–h) Quantitative analysis of the immunohistochemical staining for PCNA and caspase-3 in the ACLT rats. **P < 0.01 and ***P < 0.001. ns: not significant.

Article Snippet: Specific primary antibodies against proliferating cell nuclear antigen (PCNA) (Abclonal, 1:100), caspase-3 (Abclonal, 1:50), Col II (Abcam, 1:200), Col X (Abcam, 1:50), MMP-13 (Bioss, 1:50), SRY-related HMG box 9 (SOX9) (Abclonal, 1:50), and Runx-2 (Abcam, 1:50) were used.

Techniques: Staining, Comparison, TUNEL Assay, Immunohistochemical staining