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. 2021 Dec 13;23(2):e51287. doi: 10.15252/embr.202051287

Figure EV4. RASSF1A induction confers a differentiation phenotype accompanied by HES1 inactivation in cancer cells.

Figure EV4

  1. H1299 cells transfected with either pCDNA3 or FLAG‐RASSF1A and subjected to Matrigel 2D differentiation assay. RASSF1A expression induces differentiation as indicated by changes in the round shape and reduction of stem cell markers assessed by immunofluorescence.
  2. Quantification of NANOG and SOX2 signal intensities from (A).
  3. H1299 cells transfected with either pCDNA3 or FLAG‐RASSF1A were lysed and probed with the indicated antibodies confirming a reciprocal HES1/RNF4 regulation in response to RASSF1A.
  4. HeLa cells transfected with either siCTRL or siRASSF1A were subjected or not to treatment with the GSI DBZ (100 nM) and their lysates were immunoblotted against indicated antibodies. Cleaved Notch (ICN1) levels were used as readout of DBZ treatment.
  5. qPCR for HES1 mRNA levels in ESC in response to the γ‐secretase inhibitor DBZ (100 nM) or the Notch ligand DLL4 (1 μg/ml).

Data information: Scale bars: 50 μm. ***P < 0.001, of Student’s t‐test. Error bars indicate s.e.m. Data shown are representative of three biological replicates (n = 3).

Source data are available online for this figure.