Journal: Developmental Cell

Article Title: RhoD Inhibits RhoC-ROCK-Dependent Cell Contraction via PAK6

doi: 10.1016/j.devcel.2017.04.010

Figure Lengend Snippet: Pak6 Inhibits RhoC-Driven Cell Contraction (A) Quantification of the area of HeLa cells treated for 72 hr with the indicated siRNA and infected with the ΔF11L virus for 3 hr 40 min. (B) Images showing the subcellular localization of GFP-tagged Pak4, Pak5, and Pak6 during WR infection. Only GFP-Pak6 is associated with the plasma membrane, and mCherry provides a volume marker (see ). Scale bar, 5 μm. (C) Quantification of the area of WR- or ΔF11L-infected HeLa cells treated with Pak6 siRNA and expressing GFP-tagged WT or kinase-dead (KD) Pak6. (D) The area of Pak6 depleted HeLa cells infected with ΔF11L at 3 hr 40 min post infection and treated with H1152, C3, or RhoC siRNA. (E) The area of WR-infected HeLa cells depleted of RhoD or Pak6 and expressing GFP-tagged Pak6 or RhoD, respectively. Error bars in graphs represent the SEM from three independent experiments, in which a total of 90 (A and D) or 60 (C and E) cells were analyzed. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001; ns, not significant. See also Figure S6 .

Article Snippet: Rabbit Polyclonal Anti-Human PAK5 , ProSci , Cat#3075; RRID: AB_735027.

Techniques: Infection, Marker, Expressing

Journal: Developmental Cell

Article Title: RhoD Inhibits RhoC-ROCK-Dependent Cell Contraction via PAK6

doi: 10.1016/j.devcel.2017.04.010

Figure Lengend Snippet:

Article Snippet: Rabbit Polyclonal Anti-Human PAK5 , ProSci , Cat#3075; RRID: AB_735027.

Techniques: Transduction, Western Blot, Recombinant, Protease Inhibitor, Transfection, Software, Microscopy, Imaging

Journal: eLife

Article Title: mTORC1-induced retinal progenitor cell overproliferation leads to accelerated mitotic aging and degeneration of descendent Müller glia

doi: 10.7554/eLife.70079

Figure Lengend Snippet: ( A ) Telomere lengths of the retinal cells, which were isolated from P30 Tsc1 fl/+ and Tsc1 fl/fl mice with the indicated Cre drivers, were compared with that of control sample, and relative values are shown in the graph. Error bars denote SD. Numbers of samples analyzed are shown in the graph. All samples were obtained from independent litters. **p < 0.01; n.s., not significant. ( B ) Telomere lengths of the FACS-isolated retinal cells from P14 Tsc1 fl/+ ;Tyrp1-Cre and Tsc1 fl/fl ;Tyrp1-Cre littermate mice were compared with that of control sample, and their relative values are shown in the graph. *p < 0.05; ***p < 0.001. ( C ) Senescence markers (β-gal, p53, p21, and c-Myc) expressed in the corresponding P30 Tsc1 fl/+ ;Cre and Tsc1 fl/fl ;Cre mouse retinas were detected by Western blot (WB). Relative amounts of the proteins used in each sample were determined by WB detection of β-actin. ( D ) Expression of senescence markers (β-gal and p53) in the MG was determined by co-immunostaining with an MG marker, Sox9. The cells experienced Cre-mediated recombination were also visualized by R26tdTom reporter. Images in the right columns are the magnified versions of the boxed areas indicated by corresponding numbers in the low magnification images. ( E ) Sox9-positive MG, Pax6-positive amacrine cell (AC), and Vsx2-positive bipolar cell (BC) populations that express the senescence markers were determined and shown in the graph (Pax6 and Vsx2 staining images are provided in ). Number of samples analyzed are shown in the graph (five independent litters). n.d., not detected. ( F ) Eye and retinal structures of P30 Rptor fl/+ ;Tyrp1-Cre and Rptor fl/fl ;Tyrp1-Cre mice were investigated by hematoxylin and eosin (H&E) staining of the eye sections. Distribution of the Cre-affected cells and mTORC1 activation of the cells were examined by the immunostaining of R26tdTom and pS6, respectively. ( G ) R26tdTom-positive Cre-affected cell population in the mouse retinas were quantified by FACS and shown in the graph. ****p < 0.0001. ( H ) Telomere lengths of unsorted (whole retina) and the FACS-isolated retinal cells from P14 Rptor fl/+ ;Tyrp1-Cre and Rptor fl/fl ;Tyrp1-Cre mice were compared with that of control sample and their relative values are shown in the graph. Number of samples analyzed are shown in the graph. *p < 0.05. ( I ) Relative levels of senescence markers and mTORC1 pathway components of the mouse retinas were analyzed by WB. ( J ) Distribution of the cells expressing senescence markers and R26tdTom Cre reporter in the retinas was also examined by immunostaining.

Article Snippet: Antibody , Anti-p21/Cip1(Mouse monoclonal) , Santa Cruz Biotechnology , SC817 , WB (1:1000)IHC (1:200).

Techniques: Isolation, Western Blot, Expressing, Immunostaining, Marker, Staining, Activation Assay

Journal: eLife

Article Title: mTORC1-induced retinal progenitor cell overproliferation leads to accelerated mitotic aging and degeneration of descendent Müller glia

doi: 10.7554/eLife.70079

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-p21/Cip1(Mouse monoclonal) , Santa Cruz Biotechnology , SC817 , WB (1:1000)IHC (1:200).

Techniques: Imaging, Protease Inhibitor, Recombinant, ATP Assay, In Situ, TUNEL Assay, Software, Sequencing, CRISPR