Journal: Molecules

Article Title: Proteome Analysis of Aflibercept Intervention in Experimental Central Retinal Vein Occlusion

doi: 10.3390/molecules27113360

Figure Lengend Snippet: Proteins that significantly changed in content following aflibercept intervention.

Article Snippet: After heating, slides remained in buffer for 15 min. Primary antibodies used for immunohistochemistry included a polyclonal rabbit anti-fibronectin antibody (A0245, Dako, Glostrup, Denmark) 1:1000, a primary polyclonal rabbit anti-galectin-3 antibody (MBS3211803, MyBioSource, San Diego, CA, USA) 1:200, a monoclonal mouse anti-neurofilament light polypeptide (M0762, Dako, Glostrup, Denmark) 1:100, a rat monoclonal anti-endoplasmin antibody (MBS439463, MyBioSource, San Diego, CA, USA) and a rabbit polyclonal anti-pigment epithelium derived factor (PEDF) antibody (MBS2027143, MyBioSource, San Diego, CA, USA).

Techniques:

Journal: Molecules

Article Title: Proteome Analysis of Aflibercept Intervention in Experimental Central Retinal Vein Occlusion

doi: 10.3390/molecules27113360

Figure Lengend Snippet: Volcano plot. Log 2 of the ratio aflibercept/NaCl is plotted on the x-axis. On the y-axis, −log p -value refers to the logarithmized p -value from the t -test used to test if a protein was significantly changed. Statistically significantly changed proteins are located above the horizontal line, which denotes a significance level of 0.05. Components of aflibercept are not included in the volcano plot. PEDF: pigment epithelium-derived factor. DNAJ7C: DnaJ homolog subfamily C member 7. AKAP8: A-kinase anchor protein 8. XAB2: Pre-mRNA-splicing factor SYF1. MRPS7: 28S ribosomal protein S7, mitochondrial. RPS18: 40S ribosomal protein S18. RER1: Protein RER1.

Article Snippet: After heating, slides remained in buffer for 15 min. Primary antibodies used for immunohistochemistry included a polyclonal rabbit anti-fibronectin antibody (A0245, Dako, Glostrup, Denmark) 1:1000, a primary polyclonal rabbit anti-galectin-3 antibody (MBS3211803, MyBioSource, San Diego, CA, USA) 1:200, a monoclonal mouse anti-neurofilament light polypeptide (M0762, Dako, Glostrup, Denmark) 1:100, a rat monoclonal anti-endoplasmin antibody (MBS439463, MyBioSource, San Diego, CA, USA) and a rabbit polyclonal anti-pigment epithelium derived factor (PEDF) antibody (MBS2027143, MyBioSource, San Diego, CA, USA).

Techniques: Derivative Assay

Journal: Molecules

Article Title: Proteome Analysis of Aflibercept Intervention in Experimental Central Retinal Vein Occlusion

doi: 10.3390/molecules27113360

Figure Lengend Snippet: Immunohistochemistry comparing CRVO + aflibercept vs. CRVO + NaCl. Proteome analysis showed a small downregulation of endoplasmin and PEDF, which was further explored with immunohistochemistry. Immunohistochemistry showed similar staining patterns of ( A – D ) endoplasmin and ( E – H ) PEDF regardless of aflibercept intervention. Thus, immunohistochemistry of retinas treated with aflibercept did not show major changes in regulation of the endoplasmin or PEDF in specific retinal layers. Scale bar = 100 µm. Reaction color: brown. Abbreviations: NFL: nerve fiber layer; GCL: ganglion cell layer; IPL: inner plexiform layer; INL: inner nuclear layer; OPL: outer plexiform layer; ONL: outer nuclear layer; PS: photoreceptor segments.

Article Snippet: After heating, slides remained in buffer for 15 min. Primary antibodies used for immunohistochemistry included a polyclonal rabbit anti-fibronectin antibody (A0245, Dako, Glostrup, Denmark) 1:1000, a primary polyclonal rabbit anti-galectin-3 antibody (MBS3211803, MyBioSource, San Diego, CA, USA) 1:200, a monoclonal mouse anti-neurofilament light polypeptide (M0762, Dako, Glostrup, Denmark) 1:100, a rat monoclonal anti-endoplasmin antibody (MBS439463, MyBioSource, San Diego, CA, USA) and a rabbit polyclonal anti-pigment epithelium derived factor (PEDF) antibody (MBS2027143, MyBioSource, San Diego, CA, USA).

Techniques: Immunohistochemistry, Staining

Journal: Molecules

Article Title: Proteome Analysis of Aflibercept Intervention in Experimental Central Retinal Vein Occlusion

doi: 10.3390/molecules27113360

Figure Lengend Snippet: Western blots comparing CRVO + Eylea vs. CRVO + NaCl. ( A – B ) Western blotting was performed in two replicates. ( C – D ) Averages of quantitative data from the two replicates. The changes observed by mass spectrometry in endoplasmin and PEDF following aflibercept intervention were too slight to be confirmed by Western blotting. ( E – G ) Horizontal lines of the plots denote the means of the quantitative data. Standard deviations were larger in data obtained with Western blotting compared with quantitative data obtained through proteomic analysis. Standard deviations of endoplasmin quantification with mass spectrometry and Western blotting were 0.14 and 0.43, respectively. Standard deviations of PEDF quantification with mass spectrometry and Western blotting were 0.23 and 0.76, respectively.

Article Snippet: After heating, slides remained in buffer for 15 min. Primary antibodies used for immunohistochemistry included a polyclonal rabbit anti-fibronectin antibody (A0245, Dako, Glostrup, Denmark) 1:1000, a primary polyclonal rabbit anti-galectin-3 antibody (MBS3211803, MyBioSource, San Diego, CA, USA) 1:200, a monoclonal mouse anti-neurofilament light polypeptide (M0762, Dako, Glostrup, Denmark) 1:100, a rat monoclonal anti-endoplasmin antibody (MBS439463, MyBioSource, San Diego, CA, USA) and a rabbit polyclonal anti-pigment epithelium derived factor (PEDF) antibody (MBS2027143, MyBioSource, San Diego, CA, USA).

Techniques: Western Blot, Mass Spectrometry