Journal: Cell Proliferation

Article Title: Proliferation of myofibroblasts in the stroma of renal oncocytoma

doi: 10.1111/j.1365-2184.2010.00681.x

Figure Lengend Snippet: Protocols for immunostaining

Article Snippet: To reduce non‐specific background staining, sections were pre‐incubated with either (i) normal rabbit (X0902; Dako, Glostrup, Denmark) or swine serum (X0901; Dako, Glostrup, Denmark) at dilution of 1:25 for 15 min, or (ii) 3% bovine serum albumin (Sigma, St Louis, MO, USA) at dilution of 1:25 for 15 min. After washing in PBS, sections were incubated for 35 min with primary antibodies against: (i) MIB‐1 (M7240; Dako, Glostrup, Denmark) at 1:100 dilution, (ii) active caspase 3 (AF835; R&D, Minneapolis, MN, USA) at 1:100 dilution, (iii) SMA (A2547; Sigma) 1:2000 dilution, (iv) β‐catenin (610154; BD, San Jose, CA, USA) 1:100 dilution, (v) E‐cadherin (SC8426; Santa‐Cruz, CA, USA) 1:30 dilution, (vi) collagen I (1310‐01; Southern Biotech, Birmingham, AL, USA) 1:300 dilution, (vii) collagen III (ab6310; ABcam, Cambridge, UK) 1:600 dilution, (viii) fibronectin (ab2413; ABcam, Cambridge, UK) 1:300 dilution, (ix) desmin (M0760; Dako, Glostrup, Denmark) 1:50 dilution and (x) human caldesmon (M3557; Dako, Glostrup, Denmark) 1:200 dilution.

Techniques: Plasmid Preparation

Journal: Cell Proliferation

Article Title: Proliferation of myofibroblasts in the stroma of renal oncocytoma

doi: 10.1111/j.1365-2184.2010.00681.x

Figure Lengend Snippet: Analysis of tumour stroma of oncocytomas of the kidney. Renal oncocytomas were composed of two independent compartments, benign oncocytes and a pronounced fibrotic stroma (desmoplasia) and were associated with excessive deposition of extracellular matrix, which was periodic acid Schiff‐positive (a, pink, asterisk), collagen I‐positive (b, brown, asterisk), fibronectin‐positive (c, brown, asterisk) and collagen III‐positive (d, brown, asterisk), but negative for desmin (e) and human caldesmon (g) immunostaining. Archival smooth muscle tissue was used as positive control for desmin (f, brown, asterisk) and human caldesmon immunostaining (h, brown, asterisk) (magnification, ×400).

Article Snippet: To reduce non‐specific background staining, sections were pre‐incubated with either (i) normal rabbit (X0902; Dako, Glostrup, Denmark) or swine serum (X0901; Dako, Glostrup, Denmark) at dilution of 1:25 for 15 min, or (ii) 3% bovine serum albumin (Sigma, St Louis, MO, USA) at dilution of 1:25 for 15 min. After washing in PBS, sections were incubated for 35 min with primary antibodies against: (i) MIB‐1 (M7240; Dako, Glostrup, Denmark) at 1:100 dilution, (ii) active caspase 3 (AF835; R&D, Minneapolis, MN, USA) at 1:100 dilution, (iii) SMA (A2547; Sigma) 1:2000 dilution, (iv) β‐catenin (610154; BD, San Jose, CA, USA) 1:100 dilution, (v) E‐cadherin (SC8426; Santa‐Cruz, CA, USA) 1:30 dilution, (vi) collagen I (1310‐01; Southern Biotech, Birmingham, AL, USA) 1:300 dilution, (vii) collagen III (ab6310; ABcam, Cambridge, UK) 1:600 dilution, (viii) fibronectin (ab2413; ABcam, Cambridge, UK) 1:300 dilution, (ix) desmin (M0760; Dako, Glostrup, Denmark) 1:50 dilution and (x) human caldesmon (M3557; Dako, Glostrup, Denmark) 1:200 dilution.

Techniques: Immunostaining, Positive Control

Journal: Cancer Medicine

Article Title: The expression and role of SUZ12 in lung adenocarcinoma

doi: 10.1002/cam4.70190

Figure Lengend Snippet: The effect of SUZ12 on metastasis related genes expression was tested by qRT‐PCR and western blotting. sh‐SUZ12 increased MMP1/2/3/9 mRNA expression (A) and TIMP1/2 protein expression (B), while decreased MMP14 mRNA expression (A), MMP1/9/14 (B), TIMP3 (B), and ITGB1/5(F) protein expression, without significantly effected ITGBs mRNA expression (E). oe‐SUZ12 increased MMP14 mRNA (C) and protein (D) expression, while decreased TIMP2 mRNA expression (C) and TIMP1/2 (D) protein expression.

Article Snippet: The list of primary antibodies: SUZ12 (1 μg/mL, Abcam Cambridge, cat no: ab12073), CDK2 (1:1000; Proteintech, USA, cat. no. 10122‐1‐AP), CDK3 (1:2000; Proteintech, USA, cat. no. 55103‐1‐AP), CDK6 (1:2000; Proteintech, USA, cat. no. 14052‐1‐AP), cyclin D1 (1:5000; Proteintech, USA, cat. no. 26939‐1‐AP), cyclin E1 (1:1000; Proteintech, USA, cat. no. 11554‐1‐AP), p18 (1:1000, BOSTER China, cat. no. M03299‐1), p19 (1:1000, BOSTER China, cat. no. MA1075), p53 (1:5000; Proteintech, USA, cat. no. 60283‐2‐Ig), p‐p53 (1:2000; Proteintech, USA, cat. no. 28961‐1‐AP), p57 (1:1000, BOSTER China, cat. no. BM4129), Rb (1:1000, BOSTER China, cat. no. BM4500), pRb (1:1000, BOSTER China, cat. no. BM4338), Bcl‐2 (1:1000; Proteintech, USA, cat. no. 26593‐1‐AP), Bax (1:2000; Proteintech, USA, cat. no. 50599‐2‐lg), E‐cadherin (1:5000; Proteintech, USA, cat. no. 20874‐1‐AP), N‐cadherin (1:3000; Proteintech, USA, cat. no. 22018‐1‐AP), vimentin (1:4000; Proteintech, USA, cat. no. 10366‐1‐AP), MMP1 (1:1000, BOSTER China, cat. no. A00733‐1), MMP2 (1:500, BOSTER China, cat. no. BM4075), MMP9 (1:1000, BOSTER China, cat. no. PB0709), MMP14 (1:1000, BOSTER China, cat. no. BM4119), TIMP1 (1:1000, Bioss China, cat. no. bs‐0415R), TIMP2 (1:1000, Bioss China, cat. no. bs‐10395R), TIMP3 (1:1000; Proteintech, USA, cat. no. 10858‐1‐AP), ITGB1 (1:1000, BOSTER China, cat. no. BM4308), ITGB3 (1:1000, BOSTER China, cat. no. BA1670), ITGB5 (1:1000, BOSTER China, cat. no. A04201‐1), nm23 (1:1000, BOSTER China, cat. no. BA3787), PD‐L1 (1:3000; Proteintech, USA, cat. no. 66248‐1‐Ig), and β‐actin (1:5000; Proteintech, USA, cat. no. 66009‐1‐Ig).

Techniques: Expressing, Quantitative RT-PCR, Western Blot