Fig. 6.
Xenograft establishes that MK2KD attenuates tumor progression. a Histopathological examination revealed less differentiated and more aggressive tumors in CAL27-MK2WT than CAL27-MK2KD. The images have been captured at 100x and the scale bar denotes 100 μm. b IHC showed expression and activation status of RBPs is more prevalent in CAL27-MK2WT as compared to CAL27-MK2KD group. The images have been captured at 880x (40x objective) and the scale bar denotes 100 μm. c Protein expression analysis using tumor lysates showed that the expression of p38, MK2 and RBPs is higher in CAL27-MK2WT as compared to CAL27-MK2KD. β tubulin served as a loading control. The graphs represent change in protein expression/activation calculated as a ratio (arbitrary units). The results are expressed as means±standard errors of the mean, n = 3. Significant differences between CAL27-MK2WT and CAL27-MK2KD groups are indicated by different alphabets (p < 0.05). d qRT-PCR analysis showed that VEGF, TNF-α and MKP1 transcripts were upregulated while p27 was downregulated in CAL27-MK2KD as compared to CAL27-MK2KD tumors (as evaluated by SYBR and TaqMan chemistry). Relative gene expression was obtained after normalization with endogenous human GAPDH and determination of the difference in threshold cycle (Ct) between CAL27-MK2WT and CAL27-MK2KD groups was performed using the 2-ΔΔCt method. All the qRT-PCR assays were performed in triplicate. The results are expressed as means±standard errors of the mean. ***, p < 0.001 represent the statistical significance compared with control