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. 2026 Mar 26;11(4):e00596-25. doi: 10.1128/msphere.00596-25

Fig 2.

IBComp-eGFPPHIT cells exhibit gene expression controlled by tet, van, and cym inducers. Western and Northern analyses show POLIB expression patterns correlate with growth rates. POLIB depletion reduces proliferation while complementation restores growth.

Independent gene expression in IBComp-eGFPPHIT cells. (A) Western blot detection of POLIBrec-PTP, eGFP, and EF1α1 from IBComp-eGFPPHIT cells independently induced with Tet (4 µg/mL), Van (250 µM), and Cym (3 µg/mL) over an 8-day induction. SE, single expressor cell line representing allelically tagged POLIB-PTP (4 × 106 cell equivalents). A total of 2 × 106 cell equivalents were loaded per lane. (B) Northern blot of total RNA from IBComp-eGFPPHIT cells. U, uninduced; Cym, induced with 3 µg/mL Cym; Tet, induced with 4 µg/mL Tet; Van, induced with 250 µM Van; VaT, induced with Van and Tet; all conditions induced for 48 h. Top, probing of POLIB mRNA. Bottom, probing of tubulin mRNA as a loading control. (C) IBComp-eGFPPHIT was grown in the absence or presence of Tet (T-4 µg/mL), Van (V-250 µM), Cym (C-3 µg/mL), and Tet and Van combined (VT). Error bars represent ± s.d. of the mean from three biological replicates. (D) Western blot detection of POLIBrec-PTP and loading control EF1α1 from IBComp-eGFPPHIT grown in 250 µM Van and 4 µg/mL Tet for 8 days. SE, single expressor allelically tagged POLIB-PTP control. A total of 2 × 106 cell equivalents were loaded per lane.