Pacsin 2 RNAi in T24 cells induces cell junctions enriched with N-cadherin. An immunofluorescence microscopic image of a cluster of pacsin 2-depleted cells stained for N-cadherin (green) and F-actin (magenta) (left), and an electron microscopic image of the junctional region between the clustering ... More about this image found in Pacsin 2 RNAi in T24 cells induces cell junctions enriched with N-cadherin....

Fig. 1. Pacsin 2 localises to the cell periphery in T24 cells. (A) Immunoblot analyses of endogenous pacsin 1, pacsin 2, pacsin 3 and α-tubulin (arrowheads) in T24 cells. Blots shown are representative of three independent experiments. (B) Localization of endogenous pacsin 1, pacsin 2 and pacsin... More about this image found in Pacsin 2 localises to the cell periphery in T24 cells. (A) Immunoblot anal...

Fig. 2. Pacsin 2 depletion induces directional cell migration in T24 cells. (A) Differential interference contrast microscopy images of migrating cells in the wound healing assay. Representative micrographs show either control RNAi cells (siCtrl) or pacsin 2 RNAi cells (siPacsin 2 #1, #2 and #3)... More about this image found in Pacsin 2 depletion induces directional cell migration in T24 cells. (A) Di...

Fig. 3. Pacsin 2 depletion induces formation of N-cadherin-rich cell–cell contacts in T24 cells. (A) Immunofluorescence micrographs of control RNAi cells (siCtrl) and pacsin 2 RNAi cells (siPacsin 2 #1, #2 and #3) stained for F-actin. Merged images show F-actin (red) with DNA (blue). Scale bar: ... More about this image found in Pacsin 2 depletion induces formation of N-cadherin-rich cell–cell contacts ...

Fig. 4. Pacsin 2-depleted cells form interdigitating membrane protrusions at cell–cell contact sites in T24 cells. Transmission electron microscopy images of cell–cell contact sites in control RNAi T24 cells (siCtrl) and pacsin 2 RNAi T24 cells (siPacsin 2) at different magnifications (700×, lef... More about this image found in Pacsin 2-depleted cells form interdigitating membrane protrusions at cell–c...

Fig. 5. Pacsin 2 is required for the internalization of N-cadherin. (A) Representative immunoblots of N-cadherin internalization experiments. Surface N-cadherin in control RNAi T24 cells (siCtrl) and pacsin 2 RNAi T24 cells (siPacsin 2) was biotinylated at 4°C, and cells were subsequently incuba... More about this image found in Pacsin 2 is required for the internalization of N-cadherin. (A) Representa...

Fig. 6. Pacsin 2 SH3 domain interacts with the N-cadherin cytoplasmic region to regulate cell–cell contact formation. (A) Schematically illustrated domain structures of human pacsin 2 and human N-cadherin. Cyto, cytoplasmic domain; EC, extracellular domain; TM, transmembrane domain. Numbers indi... More about this image found in Pacsin 2 SH3 domain interacts with the N-cadherin cytoplasmic region to reg...

Fig. 7. A N-cadherin mutant with defective pacsin 2 binding shows attenuated internalization. (A) Representative immunoblots of N-cadherin internalization experiments. Surface GFP-tagged N-cadherin was biotinylated at 4°C, and cells were subsequently incubated at 37°C for the indicated periods t... More about this image found in A N-cadherin mutant with defective pacsin 2 binding shows attenuated intern...

Fig. 8. Depletion of pacsin 2 induces an elevated number of FAs in T24 cells. (A) Immunofluorescence micrographs of control RNAi cells (siCtrl) and pacsin 2 RNAi cells (siPacsin 2 #1, #2 and #3) stained for the FA marker paxillin and F-actin. Merged images show paxillin (green), F-actin (red) an... More about this image found in Depletion of pacsin 2 induces an elevated number of FAs in T24 cells. (A) ...

Fig. 1. Intracellular pH is heterogeneous in normal and cancerous lung cell lines and median pHi significantly increases in cancer cells. (A) Schematic of single-cell pHi measurements using a stably expressed pH biosensor, mCherry–pHluorin (mCh-pHl), and the protonophore nigericin to standardize... More about this image found in Intracellular pH is heterogeneous in normal and cancerous lung cell lines a...

Fig. 2. Intracellular pH is dynamic following G1 synchronization and correlates with cyclin levels. (A) Schematic of image acquisition after palbociclib synchronization. (B) Representative images of H1299-mCh-pHl cells at indicated time points after release. Ratiometric display of pHluorin/mCher... More about this image found in Intracellular pH is dynamic following G1 synchronization and correlates wit...

Fig. 3. Intracellular pH is dynamic after release from early S phase in H1299-mCh-pHl cells and correlates with cyclin levels. (A) Schematic of image acquisition after a double-thymidine synchronization. (B) Representative images of H1299-mCh-pHl cells at indicated time points after release. Rat... More about this image found in Intracellular pH is dynamic after release from early S phase in H1299-mCh-p...

Fig. 4. Intracellular pH increases leading to G2/M, followed by rapid acidification prior to division and pHi recovery in daughter cells. (A) Representative stills from Movie 1 of a dividing H1299-mCh-pHl cell at indicated time (h). Top is Hoechst 33342 dye (DNA, cyan) and DIC merge. Bottom is... More about this image found in Intracellular pH increases leading to G2/M, followed by rapid acidification...

Fig. 5. Cells released from S phase synchronization show pHi increases, leading to G2/M, rapid acidification prior to division and pHi recovery of daughter cells. (A) Representative stills from Movie 3 of a dividing H1299-mCh-pHl cell at indicated time (h). Top is Hoechst 33342 dye (DNA, cyan)... More about this image found in Cells released from S phase synchronization show pHi increases, leading to ...

Fig. 6. Single-cell pHi manipulation shows that pHi dynamics are key regulators of the cell cycle. (A) Median plots of single-cell ΔpHi from synchronizations and asynchronous (Asynch.) time-lapses. Data reproduced from Figs 2I , 3 I, and 4 C (thymidine, normalized to 4 h, n =4; palbociclib, ... More about this image found in Single-cell pHi manipulation shows that pHi dynamics are key regulators of ...

Fig. 7. Single-cell FUCCI traces show low pHi is a cue for G1 exit, S phase requires high and low pHi, and S/G2 requires high pHi. (A) Schematic of PIP–mVenus (green) and mCherry–Geminin (magenta) fluorescence intensities during cell cycle phases. (B–D) Traces from single H1299-FUCCI cells treat... More about this image found in Single-cell FUCCI traces show low pHi is a cue for G1 exit, S phase require...