Figure 6. NCBP2-AS2 regulates the levels of CAF-derived VEGFA.
(A,B) Volcano plot showing the SILAC ratio of the proteins quantified in the total proteome (A) and secretome (B) of hypoxic cCAFs transfected with siCTL and siNCBP2-AS2. The dashed lines show the regulation cut-off of at least 1-fold change and p-value < 0.05. Results from 3 biological replicates. (C) ELISA of VEGFA in the CM from hypoxic CAFs silenced or not for NCBP2-AS2 and normoxic CAFs siCTL. One-tailed t-test. N = biological replicates. (D) RT-qPCR showing remaining mRNA levels of NCBP2-AS2 normalized to TBP in cells used in (C). (E) RT-qPCR for VEGFA in hypoxic cCAFs and pCAFs transfected with siCTL or siNCBP2-AS2. The mRNA levels were normalized to TBP. One-sample t-test. N = biological replicates. (F) RT-qPCR for NCBP2-AS2 showing the silencing efficiency in cells used in (E). NCBP2-AS2 mRNA levels were normalized to TBP. N = biological replicates. (G) Influence of NCBP2-AS2 silencing on the angiogenic secretome of hypoxic CAFs. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p<0.0001.