Journal: Journal of Cancer

Article Title: Overexpression of Peroxiredoxin 6 (PRDX6) Promotes the Aggressive Phenotypes of Esophageal Squamous Cell Carcinoma

doi: 10.7150/jca.26041

Figure Lengend Snippet: PRDX6 and Erk1/2 pathway are mutually regulated. (A) Eca-109 cells were mock infected or infected with Ad-RFP or Ad-PRDX6. The expression of p-Erk1/2 and Erk1/2 were subjected to Western blotting. Relative expression of p-Erk1/2 in each group was quantified by Image J software. (B) Eca-109 cells were mock infected or infected with lentivirus expressing shRNA-NC, sh-PRDX6-2, sh-PRDX6-3. The expression of p-Erk1/2 and Erk1/2 were subjected to Western blotting. (C) Eca-109 cells were incubated with at concentrations of 0.1, 1.0, 5.0 or 10 μM PD0325901 (Erk1/2 pathway inhibitor). The expression of p-Erk1/2 and Erk1/2 were detected by Western blotting. (D) After treatment with various concentration of PD0325901, the expression of PRDX6 was measured by Western blotting in Eca-109 cells. Data are presented as mean ± SEM and were normalized to the control cells, * P < 0.05; ** P < 0.01.

Article Snippet: The MEK inhibitor PD0325901 was purchased from Cell Signaling Technology (Beverly, MA).

Techniques: Infection, Expressing, Western Blot, Software, shRNA, Incubation, Concentration Assay, Control

Journal: eLife

Article Title: Retrograde ERK activation waves drive base-to-apex multicellular flow in murine cochlear duct morphogenesis

doi: 10.7554/eLife.61092

Figure Lengend Snippet: ( A ) 3D ERK activity map in the cochlear duct cultured ex vivo for 1 day from E12.5. ( A’ ) Cross-sectional view (medial–lateral and roof–floor plane) of ( A ). Orange dotted line indicates the floor plane shown in ( B ) and ( C ). Scale bar, 50 µm. ( B ) Time-lapse snapshots of ERK activity maps in the floor plane. Time indicates the elapsed time of live imaging. Yellow arrowheads indicate the ERK activity peak. Scale bar, 100 µm. ( C ) Time-lapse snapshots of tissue flow speed obtained by particle image velocimetry in the floor plane. Scale bar, 100 µm. ( D ) Schematic diagram showing the axis, the apex–base line for kymography, and regions of interests (ROIs). ( E ) Representative kymograph of ERK activity. The horizontal axis indicates the position on the apex–base line shown in ( D ), and the vertical axis indicates the elapsed time of live imaging. Dotted lines represent oscillatory waves from the apex to the base. Scale bar, 100 µm. ( F ) ERK wave speed with mean and s.d. n = 5 from N = 3. ( G ) Time-series ERK activity rate and extension-shrinkage rate in representative three different ROIs. ( H ) Cross-correlation between the extension-shrinkage rate and ERK activity rate. n = 12. Mean ± s.d. ( I, J ) Tissue flow speed before and after the PD0325901 treatment at 1 µM, the SU5402 treatment at 30 µM ( I ), and the cyclopamine treatment at 30 µM ( J ). n = 285. Confirmed by N = 2. Two-sample t-test, p<0.001. ( K ) Time-lapse snapshots of surface-rendered ERK activity maps in the cochlear duct at E12.5. The green corners correspond to the green corner on the images shown in ( B ) and viewed from the left-bottom corner of ( B ). Circles indicate the position of ERK activity peaks, and the connecting dotted lines indicate a trace of the peak shift. The timescale is the same as in ( B ). ( L ) Schematics for the ERK activity waves and cell flow. Figure 3—source data 1. Extracellular signal-regulated kinase activity rate and extension-shrinkage rate. Figure 3—source data 2. Particle image velocimetry speed before and after the treatments with PD0325901, SU5402, and cyclopamine.

Article Snippet: The following chemicals were used: blebbistatin (FUJIFILM Wako Pure Chemical Corporation, #021-17041), cyclopamine (FUJIFILM Wako Pure Chemical Corporation, #038-19311), SU5402 (FUJIFILM Wako Pure Chemical Corporation, #197-16731), and PD0325901 (FUJIFILM Wako Pure Chemical Corporation, #162-25291).

Techniques: Activity Assay, Cell Culture, Ex Vivo, Imaging

Journal: eLife

Article Title: Retrograde ERK activation waves drive base-to-apex multicellular flow in murine cochlear duct morphogenesis

doi: 10.7554/eLife.61092

Figure Lengend Snippet: ( A ) Roof plane (orange dotted line) in the cross-sectional view of 3D ERK activity map in the cochlea at E12.5. Scale bar, 50 µm. ( B ) Time-lapse snapshots of ERK activity map in the roof plane. Time indicates the elapsed time of live imaging. Scale bar, 100 µm. ( C ) Time-lapse snapshots of tissue flow speed obtained by the particle image velocimetry in the roof plane. The multicellular flows direct toward the elongation direction around the apex tip despite winding in the region away from the tip in the roof side. Scale bar, 100 µm. ( D ) Kymograph of ERK activity for the two samples. The horizontal axis indicates the position on the apex–base line, and the vertical axis indicates the elapsed time of live imaging. Dotted lines represent the oscillatory wave trains from the apex to the base. Scale bar, 100 µm. ( E ) Time-series data of ERK activity and the tissue flow speed for elongation in the three different regions of interests. ( F, F’ ) ERK activity before and after the MEK inhibitor PD0325901 treatment at 1 µM. Scale bar, 20 µm.

Article Snippet: The following chemicals were used: blebbistatin (FUJIFILM Wako Pure Chemical Corporation, #021-17041), cyclopamine (FUJIFILM Wako Pure Chemical Corporation, #038-19311), SU5402 (FUJIFILM Wako Pure Chemical Corporation, #197-16731), and PD0325901 (FUJIFILM Wako Pure Chemical Corporation, #162-25291).

Techniques: Activity Assay, Imaging