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. 1996 Nov 26;93(24):14014–14019. doi: 10.1073/pnas.93.24.14014

Figure 5.

Figure 5

Analysis of iNOS mRNA expression by RT–PCR and Northern blot in the presence and absence of tetracyclines. (A and B) RT–PCR analysis of iNOS and β-actin mRNA expression in RAW 264.7 cells was carried out after stimulation with LPS ± doxycycline, minocycline, or hydrocortisone. Equal amounts of RNA were analyzed for iNOS and β-actin expression. “RT control” designates preparation of RT–PCRs in the absence of reverse transcriptase using the LPS-stimulated RNA as the template from RAW 264.7 cells. Data represent one of two similar and separate experiments. (C and D) Northern blot analysis of iNOS and GAPDH mRNA expression in RAW 264.7 cells was carried out after stimulation with LPS ± doxycycline, minocycline, or hydrocortisone at 16 h (C) or 4 h (D) after stimulation. The iNOS/GAPDH signal was determined and quantitated using both a PhosphoImager and a densitometer. The percent inhibition of iNOS expression was normalized with the GAPDH signal and compared with the values of the LPS-stimulated cells. Data represent one of three similar experiments.