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. 2001 May 1;107(9):1153–1161. doi: 10.1172/JCI11846

Figure 2.

Figure 2

Reduced expression of Twist mRNA and Twist protein in mutant calvaria cells. Control (Nl) and mutant (M-Tw) calvaria cells were cultured at preconfluence (PC; 2 days) or confluence (C; 4 days), and Twist mRNA and protein levels were determined. (a) Total cellular RNA was reverse transcribed and cDNA amplified by PCR to examine the expression of Twist or GAPDH transcripts followed by Southern blot analysis. (b) At confluence (4 days), equal aliquots of nuclear cell lysates were examined for Twist protein by using Western blot analysis probed with a polyclonal affinity-purified COOH-Twist Ab or a monoclonal β-actin Ab and detected by enhanced chemiluminescence. (c) Densitometric analysis of Twist mRNA levels after correction for GAPDH (mean ± SEM of three experiments, AP < 0.05 vs. Nl cells).