Fig 2: Expression of KCNQ genes in mouse and rat trachea. (A) RT-PCR of mouse KCNQ cDNA yields a single expected product size for each KCNQ channel gene. Shown are PCR products following 30 cycles of amplification. The expected amplification products are 243 (mKCNQ1), 226 (mKCNQ2), 136 (mKCNQ3), 127 (mKCNQ4), and 110 (mKCNQ5). The DNA ladder is a 50 bp increment ladder (New England Biolabs N3236). (B) Quantitative, real-time RT-PCR from pooled mouse trachea muscle RNA (panel B1) and rat trachea muscle RNA (n = 4, panel B2) of KCNQ subunit expression normalized to ß-actin expression. Gray bars in panel (B1) are from BK ß1 knockout tissues. (C) Immunostaining against KCNQ1–4, and BK ß1 (KCNMB1) proteins in rat lung sections. Color panels on the left of (CA,CB) are pseudocolor images from double immunofluorescence staining against KCNQ (middle panels) and KCNMB1 (BK ß1) (right panels). Panels (CC,CD) include a light image of tissues in the left panels, and immunostaining in right panels. Photographs are taken with the interstitial space to the left, and lumen of the airway to the right. Smooth muscle (SM) and epithelial cell (Ep) layers are indicated. Scale bars are 25 microns.

Fig 3: Transcription of the EGFP reporter in transgenic strains is testes-specific. cDNA from Tc-ß2t-EGFP#2, Tc-Rad50-EGFP#1 and Tc-Eno-EGFP#1 transgenic lines was used to perform the RT-PCR. T. castaneum RpSL32 served as a control for cDNA quality (same samples as shown in Fig. 1). Lane#M is 50 bp DNA Ladder Catalog No. (N3236S) New England Biolabs.

Fig 4: Depletion of FXR1 alters cell cycle proteins in HNSCC.(A) Relative quantity of RNAs extracted from control and FXR1 KD cells are estimated by using qRT-PCR. GAPDH serves as a control. (B) Immunoblot analysis of senescence marker proteins in both FXR1 depleted UMSCC74 and 74B cells. ß-Actin used as a loading control. (C) Relative quantity of p21 and p53 mRNAs, extracted from shcontrol and shFXR1 treated P53wt and P53mut cells, are estimated by using qRT-PCR. GAPDH serves as a control. (D) Western blot analysis of FXR1 depleted p53Wt and mutant oral cancer cells. GAPDH is used as a loading control. (E) FXR1 depleted p53Wt and mutant oral cancer cells stained with SA-ß-gal. (F) Western blot yH2AX expression in UMSCC74A and UMSCC74B cells upon FXR1 KD. (G) Protein lysates of UMSCC74B cells are subjected to RNP IP followed by qRT-PCR analysis to measure the relative quantities of RNAs in FXR1 IP compared with control IgG IP. GAPDH serves as a loading control. (H) RT-PCR of products acquired from inputs and IPs respectively from Mouse IgG as control (Lanes 1, 3) and IP of FXR1 (Lanes 2, 4) were separated and visualized by agarose gel electrophoresis. It clearly shows that p21 mRNA and TERC RNA are enriched in the IP samples, whereas p27 is not. NEB 50bp DNA ladder #N3236S (L) was loaded as a molecular marker. (*p<0.05, **p<0.005, ***p<0.0005).