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. 2021 Jun 17;24(7):102739. doi: 10.1016/j.isci.2021.102739

Figure 2.

Figure 2

The effect of MRTF or SRF downregulation on basal and serum-induced mRNA and protein expression of the components of the HEF1-AurA-HDAC6 cilium resorption pathway

(A) The promoters of HEF1, AuroraA, and HDAC6 all contain CArG boxes.

(B, E, and G) Cells were transfected with siNR, siMRTF, or siSRF as in Figure 1A, serum-deprived for 48 hr and then left untreated or exposed to 10% serum-containing medium for 3 hr. mRNA levels were determined by qPCR. Values are expressed as fold changes compared to the siNR-transfected, serum-deprived controls (n = 3 for each gene and each condition).

(C) Cells were transfected and treated as in B, and then processed for immunoblotting for the indicated proteins. A representative blot is shown.

(D, F, and H) Densitometric quantitation of protein expression for the same conditions as shown in C. Each protein was normalized to the corresponding GAPDH level and expressed as fold change compared to siNR-transfected, serum-deprived controls (n = 3). ∗p < 0.05, ∗∗p < 0.01 and ∗∗∗p < 0.001, determined by ANOVA while & and $ p < 0.05 where the fold change in corresponding two groups were determined by ratio t test (in D NR/serum-free vs. NR/serum ($) and NR/serum vs. siSRF/serum (&); in NR/serum vs. siSRF/serum ($)).