Journal: bioRxiv

Article Title: Fibroblast mechanoperception instructs pulmonary developmental and pattern specification gene expression programs

doi: 10.1101/2024.12.26.630418

Figure Lengend Snippet: (A) Western blots for HOXA5 in WT and Thy-1 KO iHLFs cultured on 5kPa PDMS hydrogel or TCP following stimulation with 20μM SRC specific inhibitor KB SRC 4 or 10μM pan-SFK inhibitor PP2 for 6hr. Quantification of HOXA5 normalized to loading control was plotted as a ratio of normalized signal relative to DMSO vehicle control. Mean ± S.D. plotted; N=3 (Thy-1 KO -5kPa and WT-1GPa) or N=4 (WT-5kPa and Thy-1 KO -1GPa) independent experiments; WT-5kPa and WT-1GPa = 1-way ANOVA with post-hoc uncorrected Fisher’s LSD test, Thy-1 KO -5kPa and Thy-1 KO -1GPa = non-parametric Kruskal-Wallis test with post-hoc uncorrected Dunn’s test. Tests chosen based on data distribution and variance. (B) Western blots for HOXA5 in WT and Thy-1 KO iHLFs cultured on 5kPa PDMS hydrogel or TCP substrates coated with αV-promoting (Fn 4G ) or α5-promoting (Fn 9*10 ) peptide fragments of the 9-10FnIII integrin binding site. Quantification of HOXA5 normalized to loading control was plotted as a ratio of normalized signal relative to full length human Fn (Fn Full ) control. Mean ± S.D. plotted; N=3 (WT-5kPa, WT-1GPa, and Thy-1 KO -5kPa) or N=4 (Thy-1 KO -1GPa) independent experiments; WT-5kPa = 1-way ANOVA with post-hoc uncorrected Fisher’s LSD test, Thy-1 KO -5kPa = Brown-Forsythe and Welch ANOVA with post-hoc unpaired t-tests with Welch’s correction, WT-1GPa and Thy-1 KO -1GPa = non-parametric Kruskal-Wallis test with post-hoc uncorrected Dunn’s test. Tests chosen based on data distribution and variance. For all statistical tests: ns = p > 0.05; p < 0.05 (*); p < 0.01 (**); p < .001 (***); p < .0001 (****).

Article Snippet: After allowing for adhesion, cells were stimulated with 1% FBS DMEM containing 20μM SRC inhibitor KB SRC 4 (Tocris) or 10μM pan-Src Family Kinase (SFK) inhibitor PP2 (Tocris) for periods of 3hr or 6hr, respectively.

Techniques: Western Blot, Cell Culture, Control, Binding Assay

Journal: International Journal of Biological Sciences

Article Title: Breast cancer-derived CAV1 promotes lung metastasis by regulating integrin α6β4 and the recruitment and polarization of tumor-associated neutrophils

doi: 10.7150/ijbs.94153

Figure Lengend Snippet: CAV1 Promotes BC Lung Metastasis by Activating the Src/FAK/α6β4 Pathway in MDA-MB-231 Cells and Simultaneously Activating Src/PI3K Signaling Downstream of α6β4 in Lung Epithelial Cells. a: CCK8 assay was used to detect the optimal acting concentration of the Src inhibitor PP2. b-e: Western blot analysis was performed to detect the expression of CAV1 and integrin Src/FAK/α6β4 signaling pathway after overexpression, knockdown, and addition of PP2. f: Western blot was used to detect the activation of integrin α6β4 downstream signaling in lung epithelial cells. g,h: IHC staining was used to detect the activation of integrin α6β4 downstream signaling PI3K/Src in mice lungs. Bar=100um. Data ware shown as mean ± SD and assessed with One-way ANOVA test. (n=3) (ns stands for non-significant difference; *p<0.05; **p<0.01; ***p<0.001).

Article Snippet: Caveolin1 Y14 phosphorylation was inhibited by the SRC family kinase inhibitor PP2, which was obtained from Abmole Bioscience Inc (Houston, USA).

Techniques: CCK-8 Assay, Concentration Assay, Western Blot, Expressing, Over Expression, Knockdown, Activation Assay, Immunohistochemistry