Journal: Nature Communications

Article Title: Synthetic aptamer mechanoreceptors enable cell-specific force sensing and temporal control via DNA circuits

doi: 10.1038/s41467-026-70765-w

Figure Lengend Snippet: a Representative brightfield and fluorescence images of HeLa cells treated with internalization inhibitors on AS1411 MP surfaces. b Corresponding quantification of mean fluorescence intensity per cell for HeLa cells. n = 70, 70, 54, 54 (left to right) cells from 3 replicates. Kruskal-Wallis test with Dunn’s multiple comparisons. c Representative images of HeLa cells on AS1411 MP surfaces showing brightfield, mechanosignal, mEGFP-paxillin, and phalloidin-stained actin. d Line profile from ( c ) shows intensity profiles of mechanosignals, mEGFP-paxillin, and actin. e Schematic showing the source of nucleolin-mediated forces. f Quantification of mean fluorescence intensity per cell following cytoskeletal inhibition. n = 72 cells from 3 replicates. Kruskal-Wallis test with Dunn’s multiple comparisons. g Representative brightfield and fluorescent images of HepG2 cells treated with internalization inhibitors on Sgc8 MP surfaces. h Corresponding quantification of mean fluorescence intensity per cell. n = 54 cells from 3 replicates. One-way ANOVA with Bonferroni post-hoc tests. i Representative images showing plasma membrane staining, mechanosignals, and brightfield of HepG2 cells incubated on Sgc8 MP surfaces. j Line profile from ( i ) reveals spatial colocalization of mechanosignals at the inner edge of membrane invaginations. k Schematic showing the source of PTK7-mediated forces. l Brightfield and fluorescence images of HepG2 cells treated with PI3K inhibitor LY294002 on Sgc8 MP surfaces. m Corresponding quantification of mean fluorescence intensity per cell for HepG2 cells. n = 54 cells from 3 replicates. Unpaired, two-tailed Student’s t-test. n Representative brightfield and fluorescence images of HeLa cells after N-WASP siRNA knockdown on AS1411 MP surfaces, and mean fluorescence intensity per cell. n = 85, 83 cells from 3 replicates for scrambled and siRNA groups. Unpaired two-tailed Student’s t-test. o Representative brightfield and fluorescence images of HepG2 cells after N-WASP siRNA knockdown on Sgc8 MP surfaces, and mean fluorescence intensity per cell. n = 45 cells from 3 replicates. Unpaired two-tailed Student’s t-test. All graphs, except ( d , j ), are presented as mean ± s.d. a.u., arbitrary units. Scale bar = 20 µm. Source data are provided as a Source Data file.

Article Snippet: N-WASP antibody (C-1, sc-271484) was purchased from Santa Cruz.

Techniques: Fluorescence, Staining, Inhibition, Clinical Proteomics, Membrane, Incubation, Two Tailed Test, Knockdown

Journal: Journal of cell science

Article Title: A role for class I p21-activated kinases in the regulation of the excitability of the actin cytoskeleton.

doi: 10.1242/jcs.263763

Figure Lengend Snippet: Fig. 4. Formation of cortactin/actin rings is PI3K dependent. (A) Still from Movie 6 of an MEF transiently overexpressing mCherry–LifeAct and PI(3,4,5)P3 marker (GFP BTK PH) following treatment with 10 μM G5555 for at least an hour. Accompanying linescan demonstrates localisation PI(3,4,5)P3 marker relative to actin cytoskeleton. (B) Still from Movie 7 of an MEF transiently overexpressing mEGFP–LifeAct and PI(4,5)P2 marker (mCherry PH domain of PLCdelta1) following treatment with 10 μM G5555 for at least an hour. Accompanying linescan demonstrates localisation of PI(4,5)P2 marker relative to actin cytoskeleton. (C) Percentage of cells producing cortactin rings in the presence and absence of 10 μM PI3K inhibitor LY294002 alongside either cotreatment with 10 μM G5555 or overexpression of GFP–PAK1Δkin. At least 100 cells analysed per condition for each of at least three biological repeats. Inhibitors were added 1 h before fixation. (D) Schematic summarising the localisation of various proteins upon addition of G5555. Scale bars: 10 μm. All error bars indicate s.d. ***P≤0.001 (ordinary one-way ANOVA followed by a post hoc Tukey’s multiple comparison test). A.U., arbitrary units.

Article Snippet: All other plasmids were acquired from Addgene: mApple LifeAct (Addgene plasmid #54747), mEGFP LifeAct (Addgene plasmid #54610), mCherry N-WASp (Addgene plasmid #55164), Myo1e mCherry (Addgene plasmid #27698), EmeraldVinculin (Addgene plasmid #54303), mCherry Arp2 (Addgene plasmid #54980), GFP BTK PH (Addgene plasmid #51463), mCherry PH domain of PLCdelta1 (Addgene plasmid #36075), GFP-PH-Tapp1 (Addgene plasmid #161985), p40PX-EYFP (Addgene plasmid #19011), Cyfip GFP (Addgene plasmid #109139).

Techniques: Marker, Over Expression, Comparison

Journal: Biomolecules

Article Title: nWASP Inhibition Increases Wound Healing via TrKb/PLCγ Signalling.

doi: 10.3390/biom13020379

Figure Lengend Snippet: Figure 6. Protein signalling changes in response to the wiskostatin treatment. (A) Changes in the total protein expression or phosphorylation sites in response to the 0.1 µM wiskostatin treatment for 1 h according to the %CFC. Protein samples that were treated with CIP at 0.6 units/µg for 1 h (B). Positive controls (C). Integrated densities as a change from the control (D). Globally normalised expression of total TrkB protein and %CFC according to protein array analysis (E). TrkB transcript expression in the HaCaT cell line (F). Protein extracts from HaCaT cells were treated with denaturation and deglycosylation reagents (G). Samples from cells treated with 0.1 µM wiskostatin/control were denatured and then deglycosylated to examine the TrkB Y816 activity (H). Western blot analysis of TrkB Y816, PLCγ1 Y1253 and Erk1/2 of HACAT cells treated with 0.1 µM wiskostatin or 10 µM 187-1 (I,J). Immunofluorescence analysis of TrkB Y816 expression in HaCaT cells serum starved for 4 h and treated with nWASP inhibitors; scale bar indicates 50 µm (K). * indicates statistical significance (* p < 0.05, ** p < 0.01,.ns: not significant).

Article Snippet: The primary antibodies used were as follows: actin (sc-16515, Insight Biotechnology, Middlesex, UK), Erk1/2 (v114A, Promega, Southampton, UK), GAPDH (sc32233, Insight Biotechnology, Middlesex, UK), PLCγ1 (sc81, Insight Biotechnology, Middlesex, UK), pPLCγ1 (sc22141, Insight Biotechnology, Middlesex, UK), nWASP (NBP1-82512, Novus Biologicals, Abingdon, UK), TrkB Y816 (ABN1381, EMD-Millipore, Watford, UK), TrkB pan (07-225, EMD-Millipore, Watford, UK) and TrkB pan (sc-136990, Insight Biotechnology, Middlesex, UK).

Techniques: Expressing, Phospho-proteomics, Control, Protein Array, Activity Assay, Western Blot

Journal: Biomolecules

Article Title: nWASP Inhibition Increases Wound Healing via TrKb/PLCγ Signalling.

doi: 10.3390/biom13020379

Figure Lengend Snippet: Figure 7. BDNF and TrkB transcript expression in the HaCaT cells and the effects of the BDNF, SOS1 and knockdown models. PCR/qPCR results for BDNF and TrkB expression in HaCaT cells treated with 0.1 µM wiskostatin (A,B); wiskostatin-treated samples at each serum starvation time point (C). qPCR analysis results of the TrkB transcript expression (D) and controls (E). Western blot analysis of TrkB Y816 (F). HaCaT cells were treated with rhBDNF at 50 and 100 ng/mL (G) and 150 ng/mL (H). Cells treated with 50 or 100 ng/mL BDNF and 0.1 µM wiskostatin (I,J). TrkB phosphorylation and SOS1 (K) and the integrated density (L). HaCaT cells treated with SOS1 (M). The effect of nWASP inhibition on TrkB Y816 phosphorylation (N). The effect of 0.1 µM wiskostatin and 10 µM 187-1 on TrkB Y816 phosphorylation (O). * indicates statistical significance (p < 0.05). ns: not significant.

Article Snippet: The primary antibodies used were as follows: actin (sc-16515, Insight Biotechnology, Middlesex, UK), Erk1/2 (v114A, Promega, Southampton, UK), GAPDH (sc32233, Insight Biotechnology, Middlesex, UK), PLCγ1 (sc81, Insight Biotechnology, Middlesex, UK), pPLCγ1 (sc22141, Insight Biotechnology, Middlesex, UK), nWASP (NBP1-82512, Novus Biologicals, Abingdon, UK), TrkB Y816 (ABN1381, EMD-Millipore, Watford, UK), TrkB pan (07-225, EMD-Millipore, Watford, UK) and TrkB pan (sc-136990, Insight Biotechnology, Middlesex, UK).

Techniques: Expressing, Knockdown, Western Blot, Phospho-proteomics, Inhibition