Additional file 1: of The involvement of mast cells in the irinotecan-induced enteric neurons loss and reactive gliosis

CPT-11 increases the number of mast cells in the small intestine. Intestinal segments (duodenum, jejunum or ileum) were stained with toluidine blue. Mast cells (black arrows) were counted in all intestinal segments. Scale bar = 50 μm. (TIF 3590 kb

Additional file 1: Figure S1. of Rheological properties of cells measured by optical tweezers

Plots of K ' and K " measured for fibroblasts, neurons, and astrocytes vs immersion angle θ for the different frequencies probed in this work. (DOCX 110 kb

MOESM1 of The availability of the embryonic TGF-β protein Nodal is dynamically regulated during glioblastoma multiforme tumorigenesis

Additional file 1: Figure S1. Nodal immunostaining changes along the development of the oncospheres. (a) Small sphere showing only cells that with a symmetrical Nodal distribution in the cytoplasm. (b) Large sphere comprised by cells with distinct Nodal distributions. (c–e) Nodal immunostaining (red) in OB1 stem cells placed at the top of oncospheres is localized to the cell membrane (green, phalloidin, asterisk). (f–h) Cells located at the lateral edge of oncospheres harboring different heights presented Nodal immunostaining symmetrically distributed in the cytoplasm of cells. (i–l) Cells directly attached to the substrate also presented a symmetrical distribution of Nodal. (e, h, k, l) Optical slices projected on the YZ axis showing a virtual reconstruction of the oncosphere

Additional file 2: of The involvement of mast cells in the irinotecan-induced enteric neurons loss and reactive gliosis

Mast cell pre-degranulation prevents CPT-11-induced increase of tryptase immunostainedcells in the duodenum and jejunum of mice. Graph represents the mean ± SEM of the number of tryptasepositive cells in the duodenum, and jejunum in ten microsc opic field per section from four mice in eachgroup. White, black and crosshatch bars represent, respectively control, CPT-11 and CPT-11+c48/80 group.#P < 0.05 versus control group. *P < 0.05 versus CPT-11 group. One-way ANOVA followed by Bonferroni. (DOC 70 kb

Tether extraction and radius measurements for neurons.

<p>(A–C) Images of the cortical (CX) neurons cytoskeleton stained for F-actin with phaloidin-FITC, in green (A), β-tubulin III, in red (B) and the merge of both images (C). (D–F) Images of the ganglionic eminence (GE) neurons cytoskeleton stained for F-actin with phaloidin-FITC, in green (D), β-tubulin III, in red (E) and the merge of both images (F). (C) and (F) also display numbers locating the 3 different regions from which tethers were extracted, (1) cell body; (2) neurite; (3) growth cone. (G) Mean values of the tether force, , extracted from both Neuron CX and Neuron GE in regions 1, 2 and 3. (H) SEM image of a typical tether extracted from Neuron GE. Scale bar is 1 µm. (I) Mean values of the tether radius, <i>R</i>, extracted from both Neuron CX and Neuron GE in regions 1, 2 and 3. Standard errors were used as error bars in (G) and (H). At least 20 different experiments were performed for each situation in (G) and (H). (J)–(L) Images of tethers extracted from Neuron GE stained for F-actin with phaloidin-FITC, in green (J), β-tubulin III, in red (K) and the merge of both images (L). Scale bar is 5 µm.</p

Tether extraction from microglial cell.

<p>(A) Image of the extracted tether in bright field with Image J shadow north processing filter applied. (B) Zoom of the white rectangle in A. Scale bar for A is 10 µm and for B is 5 µm. (C) A typical tether extraction force curve, indicating the maximum force <i>F_m</i> and the approach to <i>F</i>₀, the steady-state tether force.</p

Plasma membrane vesicles growth dynamics.

<p>(A) Time 0min, U-87 MG cells immediately after treating with the PMV solution. (B) Time 10 min, appearance of small PMVs, indicated with white arrows. (C) Time 20 min, PMVs growth. (D) Time 30 min, some PMVs reach their maximum size. Scale bar is 50 µm.</p

Tether extraction and radius measurements for macrophage cells.

<p>(A–C) Images of the control macrophage cytoskeleton stained for F-actin with phaloidin-FITC, in green (A), stained with CD68, in red (B) and both images merged (C). (D–F) Images of the macrophage+LPS cytoskeleton stained for F-actin with phaloidin-FITC, in green (C), CD68, in red (D) and both images merged (F). Scale bars for A-F are all 10 µm. (G) Mean values of the tether force, , extracted from macrophage cells in both conditions. (H) SEM image of a typical tether extracted from macrophage+LPS cells. Scale bar is 1 µm. (I) Mean values of the tether radius, <i>R</i>, extracted from macrophage cells in both conditions. Standard errors were used as error bars in (G) and (I). At least 20 different experiments were performed for each situation in (G) and (I). (*** means p<0.0001 in t-test statistics).</p

Surface tension and bending modulus values.

<p>Surface tension and bending modulus values.</p

Tether extraction and radius measurements for astrocytes and glioblastoma cells.

<p>(A-B) Images of the astrocyte cytoskeleton stained for F-actin with phaloidin-FITC, in green (A) and GFAP, in red (B). (C-D) Images of glioblastomas U-87 MG and GBM95 cytoskeleton, respectively stained for F-actin with phaloidin-FITC, in green. Scale bars for A-D are all 10 µm. (E) Mean values of the tether force, , extracted from astrocytes and glioblastomas cells. (F) Mean values of the tether radius,<i>R</i>, extracted from astrocytes and glioblastomas cells. Standard errors were used as error bars in (E) and (F). At least 20 different experiments were performed for each situation in (E) and (F). (G-I) Images of tethers extracted from astrocytes, stained for F-actin with phaloidin-FITC, in green (G) and GFAP, in red (H). (G) and (H) merged in (I). Scale bars for G-I are all 5 µm.</p