Journal: bioRxiv

Article Title: Mechanical stress in pancreatic cancer: Signaling pathway adaptation activates cytoskeletal remodeling and enhances cell migration

doi: 10.1101/2021.06.11.448065

Figure Lengend Snippet: A, MIA PaCa-2 and PANC-1 cells were pre-treated with 15 μM p38 MAPK inhibitor (SB202190), JNK inhibitor (SP600125) or equal volume of DMSO and subjected to a scratch wound healing assay under 4.0 mmHg of compression. Pictures from at least 4 different fields were taken from three biological replicates. Scale bar: 0.2 mm. White dashed line shows the difference in wound closure between 0 and 16 hours. B-C, Graph showing the average ±SE percentage wound closure of compressed MIA PaCa-2 (B) and PANC-1 (C) treated with DMSO or inhibitors from 3 biological replicates (n≥12). Asterisk (*) indicates a statistically significant difference (p<0.05 in student’s t test). D-E , Graph showing the average Ki67 area fraction in compressed MIA PaCa-2 (D) and PANC-1 (E) cells treated with DMSO, p38 MAPK or JNK inhibitor from at least 10 different fields/ condition from two biological replicates as quantified automatically using an in-house code in MATLAB. Asterisk (*) indicates a statistically significant difference in student’s t test (p<0.05). F , Representative Western Blotting showing phosphorylated HSP27 (Ser 82) and c-Jun (S73) in control and compressed MIA PaCa-2 cells treated with 15 μM of each inhibitor or equal volume of DMSO. Antibody against β-actin was used as a loading control. Quantification of each antibody compared to loading control was quantified by ImageJ and it is indicated by numbers in grey font.

Article Snippet: For image analysis, the calculation of Ki67 area fraction was performed automatically using a previously developed inhouse code in MATLAB (MathWorks, Inc., Natick,MA) , , .

Techniques: Wound Healing Assay, Western Blot, Control

Journal: bioRxiv

Article Title: Mechanical stress in pancreatic cancer: Signaling pathway adaptation activates cytoskeletal remodeling and enhances cell migration

doi: 10.1101/2021.06.11.448065

Figure Lengend Snippet: A-B , Rac1 (A) and cdc42 (B) mRNA expression was quantified by qPCR in both MIA PaCa-2 and PANC-1. Each bar indicates the mean fold change ±SE of two biological replicates (n=6). Asterisk (*) indicates a statistically significant difference (p<0.05 in student’s t test). C , MIA PaCa-2 and PANC-1 pancreatic cancer cells were treated with siRNA against Rac1 (siRac1) or cdc42 (sicdc42) and then subjected to a scratch wound healing assay for 16 hours under 0.0 or 4.0 mmHg of compression in 2 % FBS containing DMEM. Control cells were treated with stealth siRNA (siCTRL). Scale bar: 0.1 mm. D-E , Graphs showing the percentage ±SE wound closure of MIA PaCa-2 (D) and PANC-1 (E) as quantified using ImageJ software. Statistically significant difference in wound closure of compressed siRac1- or sicdc42-treated cells compared to siCTRL-treated cells is indicated with an asterisk (*) (2 biological replicates; n≥6; p<0.05 in student’s t test). F , Graph showing the average ±SE Ki67 area fraction in MIA PaCa-2 and PANC-1 control and compressed cells treated with siCTRL, siRac1 or sicdc42 from at least 5 different fields/ condition from two biological replicates as quantified automatically using an in-house code in MATLAB. No statistically significant changes were observed.

Article Snippet: For image analysis, the calculation of Ki67 area fraction was performed automatically using a previously developed inhouse code in MATLAB (MathWorks, Inc., Natick,MA) , , .

Techniques: Expressing, Wound Healing Assay, Control, Software