TABLE 1.

Characteristics of isolates from CIP-treated PAO1-mCherry-P_CD-gfp+ flow cell biofilms^e

Biofilma	Isolate	MIC (μg/ml)b	Mutation	Effect on NfxB protein
FCB1 (24 h + CIP for 24 h)	FCB 2	0.75	Ins 259A	Frameshift, no stopc
	FCB 3	0.75	Ins 259A	Frameshift, no stop
	FCB 4	0.75	Ins 259A	Frameshift, no stop
	FCB 5	0.75	Ins 259A	Frameshift, no stop
	FCB 6	0.75	Ins 259A	Frameshift, no stop
	FCB 7	0.75	Ins 259A	Frameshift, no stop
	FCB 8	0.75	Ins 259A	Frameshift, no stop
	FCB 1	0.75–1.0	Ins 259A	Frameshift, no stop
FCB2 (24 h + CIP for 24 h)	FCB 9	0.75	A600C	Stop>Cys
	FCB 10	0.75	A151C	Thr51Pro
	FCB 12	0.75	A151C	Thr51Pro
	FCB 11	0.75–1.0	A151C	Thr51Pro
	FCB 13	1.0	A151C	Thr51Pro
	FCB 14	1.0	A151C	Thr51Pro
	FCB 15	1.0	Δ(115) 1 bp	Frameshift, early stop (46 aa)
	FCB 16	1.0	A151C	Thr51Pro
FCB3d (24 h + CIP for 24 h)	FCB 17	0.38–0.5	A600C	Stop>Cys
	FCB 18	0.5	A600C	Stop>Cys
	FCB 19	0.5	A151C	Thr51Pro
	FCB 20	0.75	A600C	Stop>Cys
FCB4 (72 h + CIP for 96 h)	FCB 26	1.0	T440C	Phe147Ser
	FCB 27	1.0	Δ(164–165) 2 bp	Frameshift, no stop
	FCB 28	1.0	Δ(485–491) 7 bp	Frameshift, no stop
	FCB 29	1.0	Δ(485–491) 7 bp	Frameshift, no stop
	FCB 30	1.0	C509A	Ala170Glu
FCB5 (72 h + CIP for 96 h)	FCB 31	0.5	A600C	Stop>Cys
	FCB 34	0.5	A600C	Stop>Cys
	FCB 32	1.0	Δ252 (1 bp)	Frameshift, early stop (94 aa)
	FCB 35	1.0	Δ252 (1 bp)	Frameshift, early stop (94 aa)
	FCB 33	1.5	G381A	Trp127Stop
FCB6 (72 h + CIP for 96 h)	FCB 36	1.0	G143A	48 Ser>Asn
	FCB 37	1.0	G143A	48 Ser>Asn
	FCB 39	1.0	G143A	48 Ser>Asn
	FCB 38	1.5	G381A	Trp127stop
	FCB 40	1.5	G143A	48 Ser>Asn

^aIndependent biofilms grown for 24 h or 72 h and then treated with 0.2 μg/ml CIP for 24 h or 96 h, respectively.

^bMIC of CIP as determined by Etest.

^cStop codon not detected in the sequenced region of the nfxB amplicon.

^dCombination of two biofilms.

^eIns, insertion; aa, amino acids.