Journal: bioRxiv

Article Title: ROCK2 inhibition has a dual role in reducing ECM remodelling and cell growth, while impairing migration and invasion

doi: 10.1101/2025.07.21.666015

Figure Lengend Snippet: ROCK2 inhibition with GV101 reduces CAF-mediated remodelling of pre-existing collagen in murine and human models of TNBC, also see Figures S1 and S2 (A) Schematic of collagen contraction assays utilising CAFs isolated from end-stage MMTV-PyMT tumours. (B) Representative images of PyMT CAF matrices treated with DMSO, GV101 or Fasudil at days 3 and 12 of contraction (i) and quantification of PyMT CAF organotypic matrix size over a 12-day contraction period (ii) and on day 12 (iii). (C) Representative SHG images of PyMT CAF matrices to assess collagen I (top panel, magenta), picrosirius red staining to assess fibrillar collagen (middle panel) and polarised light birefringence imaging to assess collagen bundling and density (bottom panel). (D) Quantification of unconfined compression analysis to assess Young’s modulus. (E-G) Quantification of PyMT CAF matrices; SHG peak signal intensity (E), picrosirius red staining coverage (F) and total birefringence signal (G). (H) Schematic of collagen contraction assays utilising CAFs isolated from human TNBC patients. (I) Representative images of human CAF matrices treated with DMSO, GV101 or Fasudil at days 3 and 12 of contraction (i) and quantification of CAF organotypic matrix size over a 12-day contraction period (ii) and on day 12 (iii). (J) Representative SHG images of human CAF matrices to assess collagen I (top panel, magenta), picrosirius red staining to assess fibrillar collagen (middle panel) and polarised light birefringence imaging to assess collagen bundling and density (bottom panel). (K) Quantification of unconfined compression analysis to assess matrix Young’s modulus. (L-N) Quantification of human CAF matrices; SHG peak signal intensity (L), picrosirius red staining coverage (M) and total birefringence signal (N). Data represents mean ± SEM of three individual repeats. p-values were determined using One-sample t and Wilcoxon test for normalised data and for remaining data a One-way ANOVA with multiple comparisons was used. ns = not significant (p≥0.05), * = p<0.05, ** = p<0.01, *** = p<0.001, **** = p<0.0001. Scale bar = 1cm (B and I), 100µm (C and J).

Article Snippet: Slides were then incubated in 0.02% phosphomolybdic acid for 2 minutes and briefly washed in water before a 2 hour incubation in 0.1% Picrosirius Red solution (Australian Biostain solution).

Techniques: Inhibition, Isolation, Staining, Imaging

Journal: bioRxiv

Article Title: ROCK2 inhibition has a dual role in reducing ECM remodelling and cell growth, while impairing migration and invasion

doi: 10.1101/2025.07.21.666015

Figure Lengend Snippet: (A) Schematic of cell-derived matrix (CDM) production assays where both PyMT and human CAFs are treated with ascorbic acid to stimulate matrix production in the presence of DMSO, GV101 or Fasudil. (B) Representative images of PyMT CAF derived CDMs showing SHG imaging (top panel, magenta) and picrosirius red stained matrices (bottom panel). (C) Quantification of peak SHG signal (i) and picrosirius red staining coverage for PyMT CAF generated CDMs (ii). (D) Representative images of human CAF derived CDMs showing SHG imaging (top panel, magenta) and picrosirius red stained matrices (bottom panel). (E) Quantification of peak SHG signal (i) and picrosirius red coverage (ii) for human CAF generated CDMs. Data represents mean ± SEM of three individual repeats. p-values were determined using One-sample t and Wilcoxon test for normalised data and for remaining data a One-way ANOVA with multiple comparisons was used. ns = not significant (p≥0.05), * = p<0.05, ** = p<0.01. Scale bar = 100μm.

Article Snippet: Slides were then incubated in 0.02% phosphomolybdic acid for 2 minutes and briefly washed in water before a 2 hour incubation in 0.1% Picrosirius Red solution (Australian Biostain solution).

Techniques: Derivative Assay, Imaging, Staining, Generated

Journal: Frontiers in Cell and Developmental Biology

Article Title: Cardiac Fibroblast Growth Factor 23 Excess Does Not Induce Left Ventricular Hypertrophy in Healthy Mice

doi: 10.3389/fcell.2021.745892

Figure Lengend Snippet: Adeno-associated virus expressing murine Fgf23 mice do not show any signs of pathological LVH. (A) Representative cross-sections of AAV-Fgf23 and Ctrl mice stained with HE (original magnification ×10). (B) Representative cross-sections of AAV-Fgf23 and Ctrl mice stained and wheat germ agglutinin (WGA) (original magnification ×40; scale bar, 50 μm). (C) Quantification of at least 100 individual cardiac myocytes per mouse reveals no size differences between both groups. (D) As analyzed by quantitative real-time PCR, cardiac Fgfr4 mRNA expression is significantly enhanced in AAV-Fgf23 mice compared to Ctrl. (E) The pro-hypertrophic NFAT target genes BNP , bMHC , Rcan1 , and Trpc6 are not induced in AAV-Fgf23 mice compared to Ctrl. (F) Representative images of picrosirius red-stained mid-chamber free-wall of AAV-Fgf23 and Ctrl mice (original magnification, ×63; scale bar, 50 μm). (G,H) The mRNA and protein expression of fibrosis-associated markers Tgfb1, Col1a1, and Ctgf and are not altered in AAV-Fgf23 mice. Gapdh serves as the loading control. Data are given as scatter dot plots with means; *** p < 0.001 analyzed using Mann–Whitney test according to D’Agostino and Pearson’s normality test; n = 6–14 mice per group.

Article Snippet: For the quantification of interstitial LV fibrosis, sections were incubated using picrosirius red solution (Merck) in 1.2% picric acid and analyzed using ImageJ.

Techniques: Expressing, Staining, Real-time Polymerase Chain Reaction, MANN-WHITNEY