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. Author manuscript; available in PMC: 2019 Oct 15.
Published in final edited form as: Sci Signal. 2019 Feb 5;12(567):eaan8247. doi: 10.1126/scisignal.aan8247

Figure 7. NCBP2-AS2 regulates EC sprouting through VEGFA signaling.

Figure 7

(A) Sprouting quantification of ECs treated with the CM from hypoxic cCAFs transfected with siCTL or siNCBP2-AS2 (two different siRNAs) with or without VEGFA. N = beads assessed from 3 biological replicates. CM without VEGFA: siCTL n = 149, siNCBP2-AS2 #1 n = 128 and siNCBP2-AS2 #2 n = 110. CM with VEGFA: siCTL n=132, siNCBP2-AS2 #1 n = 130 and siNCBP2-AS2 #2 n = 126. (B) Sprouting quantification of ECs treated with the CM from hypoxic pCAFs transfected with siCTL or siNCBP2-AS2 (two different siRNAs) with or without VEGFA. N = beads assessed from 3 biological replicates. CM without VEGFA: siCTL n = 162 beads, siNCBP2-AS2 #1 n = 132 and siNCBP2-AS2 #2 n = 124. CM with VEGFA: siCTL n=127, siNCBP2-AS2 #1 n = 133 and siNCBP2-AS2 #2 n = 136. (C) RT-qPCR showing remaining mRNA levels of NCBP2-AS2 normalized to TBP. (D) Sprouting quantification of ECs treated with the CM from hypoxic cCAFs transfected with siCTL or siNCBP2-AS2 (two different siRNAs) in the presence of bevacizumab or IgGK (control). N = beads assessed from 3 biological replicates. CM with IgG: siCTL n = 135, siNCBP2-AS2 #1 n = 129 and siNCBP2-AS2 #2 n = 107. CM with bevacizumab: siCTL n = 127, siNCBP2-AS2 #1 n = 147 and siNCBP2-AS2 #2 n = 122. (E) Sprouting quantification of ECs treated with the CM from hypoxic pCAFs transfected with siCTL or siNCBP2-AS2 (two different siRNAs) in the presence of bevacizumab or IgGK (control). N = beads assessed from 3 biological replicates. CM with IgG: siCTL n = 121, siNCBP2-AS2 #1 n = 108 and siNCBP2-AS2 #2 n = 107. CM with bevacizumab: siCTL n = 126, siNCBP2-AS2 #1 n = 130 and siNCBP2-AS2 #2 n = 110. (F) RT-qPCR showing the remaining mRNA levels of NCBP2-AS2 normalized to TBP. (G) Representative western blot for proteins downstream of VEGFR signaling in ECs treated with the CM from cCAFs silenced or not for NCBP2-AS2. N = 3 biological replicates. (H) RT-qPCR showing remaining mRNA levels of NCBP2-AS2 normalized to TBP. (I) Working model. T-test. * p < 0.05, ** p < 0.01, **** p < 0.0001, NS: not significant.