Fig. 4.

Med12 knockdown increases cell adhesion and preserves cell viability on limiting (gelatin) substratum during neuronal differentiation. a-d mNS-5 NSCs infected with lentiviruses expressing NS control, Med12-, or Cdk8-specific shRNAs were seeded into T-75 tissue culture flasks coated with gelatin (a, b) or laminin (c, d) prior to initiation of neuronal differentiation as described in Fig. 3. Fluorescence based adhesion assay (a, c) was performed 40 h after initiation of the neuronal differentiation. Data represent the mean +/− SEM of at least three independent experiments performed in triplicate. p values were calculated by Student’s t test. Brightfield images (b, d) were obtained by optical microscopy at 1, 4, and 7 days after initiation of neuronal differentiation. e and f Validation of Med12 and Cdk8 depletion in knockdown cells by RT-qPCR (e) and immunoblot (f) analyses. mRNA levels for each gene in (e) were normalized to β-actin mRNA and expressed relative to their corresponding mRNA levels in untreated (MOCK) cells. Data represent the mean +/− SEM of at least three independent experiments performed in triplicate. p values were calculated by Student’s t test