Journal: Journal of cellular physiology

Article Title: Stimulation of vascular smooth muscle cell proliferation by stiff matrix via the IK Ca channel-dependent Ca 2+ signaling.

doi: 10.1002/jcp.30349

Figure Lengend Snippet: FIGURE 2 Stiff substrates upregulate the IKCa channel expression in A7r5 VSMCs. (a, b) Representative images showing the IKCa mRNA expression using RT‐PCR and IKCa protein cell surface expression using flow cytometry. (c) Summary of the mean data from 4 independent experiments. *p < .05; **p < .01. GAPDH, glyceraldehyde 3‐phosphate dehydrogenase; IKCa, intermediate‐conductance Ca2+‐activated K+; mRNA, messenger RNA; RT‐PCR, reverse‐transcription polymerase chain reaction;VSMC, vascular smooth muscle cell

Article Snippet: Following three washes with PBS, cells were incubated with a pri- mary mouse antibody specifically against the IKCa extracellular domain (ALM051, 1:100; Alomone Labs) for 40min.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Flow Cytometry, Reverse Transcription, Polymerase Chain Reaction

Journal: Journal of cellular physiology

Article Title: Stimulation of vascular smooth muscle cell proliferation by stiff matrix via the IK Ca channel-dependent Ca 2+ signaling.

doi: 10.1002/jcp.30349

Figure Lengend Snippet: FIGURE 3 Stiff substrate‐induced IKCa channel upregulation is required for increased A7r5 VSMC proliferation. (a) Summary of the effects of treatment with 100 nM TRAM34 on stiff substrate‐induced cell proliferation from three independent experiments. (b) Representative western blots showing siRNA‐mediated knockdown of the IKCa protein expression (top) and summary of the data from five independent experiments (bottom). (c) Summary of stiff substrate‐induced cell proliferation in siRNA‐transfected cells from three independent experiments. *p < .05 and **p < .01 compared between 0.21 and 1.72 MPa without any treatment or compared between 1.72 MPa with or without treatment with TRAM34, respectively. IKCa, intermediate‐conductance Ca2+‐activated K+; siRNA, small interfering RNA; VSMC, vascular smooth muscle cell

Article Snippet: Following three washes with PBS, cells were incubated with a pri- mary mouse antibody specifically against the IKCa extracellular domain (ALM051, 1:100; Alomone Labs) for 40min.

Techniques: Western Blot, Knockdown, Expressing, Transfection, Small Interfering RNA

Journal: Journal of cellular physiology

Article Title: Stimulation of vascular smooth muscle cell proliferation by stiff matrix via the IK Ca channel-dependent Ca 2+ signaling.

doi: 10.1002/jcp.30349

Figure Lengend Snippet: FIGURE 4 Stiff substrate induces IKCa‐ dependent increase in [Ca2+]i in A7r5 VSMCs. (a) Representative single‐cell images showing intracellular Ca2+ concentration in cells cultured on the soft and stiff substrate under indicated conditions. Treatment with 100 nM TRAM34 was made during culture media and also in extracellular recording solution. (b) Summary of the relative Fluo4 fluorescence intensity, indicative of the [Ca2+]i, in individual cells under the condition shown in (a), with 375 cells examined for each case. **p < .01. IKCa, intermediate‐conductance Ca2+‐activated K+; VSMC, vascular smooth muscle cell

Article Snippet: Following three washes with PBS, cells were incubated with a pri- mary mouse antibody specifically against the IKCa extracellular domain (ALM051, 1:100; Alomone Labs) for 40min.

Techniques: Concentration Assay, Cell Culture, Fluorescence

Journal: Journal of cellular physiology

Article Title: Stimulation of vascular smooth muscle cell proliferation by stiff matrix via the IK Ca channel-dependent Ca 2+ signaling.

doi: 10.1002/jcp.30349

Figure Lengend Snippet: FIGURE 5 Stiff substrate‐induced IKCa channel‐dependent increases in [Ca2+]i is essential for increased A7r5 VSMC proliferation. Summary of cell proliferation on substrates in culture media containing (a) 1 mM EGTA or (b) 10 μM BAPTA‐AM, from three independent experiments for each case. **p < .01. EGTA, ethylene glycol‐bis(β‐aminoethyl ether)‐N,N,N′,N′‐tetraacetic acid; IKCa, intermediate‐conductance Ca2+‐activated K+; VSMC, vascular smooth muscle cell

Article Snippet: Following three washes with PBS, cells were incubated with a pri- mary mouse antibody specifically against the IKCa extracellular domain (ALM051, 1:100; Alomone Labs) for 40min.

Techniques: