Fig. 2

Impact of serum deprivation on de-adhesion dynamics and expression levels of VTN and p53. a De-adhesion dynamics of serum-deprived WJ-MSCs. WJ-MSCs were treated with TrypLE and the time taken by the MSCs to contract to a rounded morphology was recorded with the help of time lapse imaging. The normalized area vs. time data were fitted to a Boltzmann sigmoid equation to determine the time constants τ1 and τ2. Delayed de-adhesion leading to greater values for both the time constants were observed. Differences in τ1 and τ2 between control and serum-deprived WJ-MSCs were significant. A total of 45 cells from 3 different biological samples were compared. b mRNA expressions of VTN as detected by qRT-PCR (n = 5). c Band density of protein expression levels of VTN was quantified relative to GAPDH and plotted (n = 3). d Representative images of Western blots. GAPDH was used as a loading control. MCF7 cells were used as negative control (n = 5), as they did not show much increase in expression under serum deprivation. Out of the five sets of experiments performed, two of them showed a small increase in VTN protein expression. e The distribution of VTN (green) and vimentin (red) was visualized by immunofluorescence. Insets show nuclei stained with DAPI. Representative images from four independent biological samples are shown (n = 4). f p53 mRNA was detected by qRT-PCR (n = 4). g Band density of protein expression levels of p53 was quantified relative to GAPDH and plotted (n = 3). Serum-deprived condition represented as no serum (NS) in the figure (*p < 0.05, ***p < 0.0001)