Journal: bioRxiv

Article Title: NOX4 inhibition promotes the remodeling of dystrophic muscle

doi: 10.1101/2022.01.08.475493

Figure Lengend Snippet: ( A ) Representative immunofluorescent images from Nox4 WT : mdx and Nox4 KO : mdx gastrocnemius (gastroc) muscles stained with anti-NOX4 antibody. Muscle fibrosis was histologically assessed in the diaphragm and gastroc muscles from 3 month-old Nox4 WT : mdx and Nox4 KO : mdx mice using picrosirius red staining (n = 4). ( B ) Representative images and ( C-D ) histological quantifications reveal no significant differences in muscle fibrosis between the groups at this age. Data are presented as box-and-whisker plots with error bars representing minimum and maximum values. Data were analyzed using unpaired, two-tailed Welch’s T-tests (α = 0.05; effect size is presented as Cohen’s d ). Scale bar represents 100 µm.

Article Snippet: Picrosirius red staining was performed as previously described ( ) following decalcification of muscle sections using Formical-2000 (StatLab).

Techniques: Muscles, Staining, Whisker Assay, Two Tailed Test

Journal: bioRxiv

Article Title: NOX4 inhibition promotes the remodeling of dystrophic muscle

doi: 10.1101/2022.01.08.475493

Figure Lengend Snippet: ( A ) Representative picrosirius red staining and image quantification of ( B ) diaphragm and ( C ) gastrocnemius muscles from 6 month-old D2.WT (n = 6), NOX4 wild-type (NOX4-WT) and knockout (NOX4-KO) littermates from a NOX4 KO line generated on the D2. mdx background (Nox4 KO : mdx ; n = 12), and D2. mdx mice that received vehicle or GK831 (GKT) treatments beginning at 3 months of age (n = 12-14). Fibrosis quantifications for 3 month-old D2. mdx mice are included to show baseline values for GKT treatment groups. Muscle function was assessed for the ( D ) diaphragm and ( E ) extensor digitorum longus (EDL) muscles of NOX4-WT (+/+) and NOX4-KO (-/-) littermates of the Nox4 KO : mdx mouse line. ( F ) Biochemical quantification of collagen content was performed on gastrocnemius muscles of vehicle and GKT-treated D2. mdx mice. Data are presented as box-and-whisker plots with error bars representing minimum and maximum values. ( D-F ) Percent values indicate the difference between mean values of the two groups. Data were analyzed using ( B-C ) ANOVA followed by Tukey post-hoc tests (α = 0.05; effect size is reported as η 2 ) or ( D-F ) unpaired, two-tailed Welch’s T-tests (α = 0.05; effect size is reported as Cohen’s d ); *p < 0.05 vs. D2.WT values; #p < 0.05 vs. 3 mo D2. mdx values; $p < 0.05 vs. respective control group values.

Article Snippet: Picrosirius red staining was performed as previously described ( ) following decalcification of muscle sections using Formical-2000 (StatLab).

Techniques: Staining, Muscles, Knock-Out, Generated, Whisker Assay, Two Tailed Test, Control

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Modulation of Cancer-Associated Fibrotic Stroma by An Integrin α v β 3 Targeting Protein for Pancreatic Cancer Treatment

doi: 10.1016/j.jcmgh.2020.08.004

Figure Lengend Snippet: ProAgio depletes CAPaSC by targeting integrin α V β 3 and resorbs tumor collagen. ( A and B ) Mean tumor volume of ( A ) subcutaneous xenograft of Panc1 with ( solid lines ) or without ( dotted lines ) co-implantation of PaSC and mean tumor weight ( B ) of Panc1 subcutaneous xenograft with or without co-implantation of PaSC upon treatment with vehicle ( black lines in panel A , grey dots in panel B ) or 10 mg/kg ProAgio (12 daily doses, red lines in panel A , red dots in panel B ) (n = 6). ( C and E ) Representative images of immunofluorescence staining of α-SMA (red) ( C ) and Masson trichrome (blue) staining for collagen ( E ). ( D and F ) Quantitative analyses of immunofluorescence staining of α-SMA ( D ) and Masson trichrome ( F ) staining in tumor sections of Panc1 xenograft (with or without PaSC co-implantation) mice treated with vehicle or 10 mg/kg ProAgio. ( G and I ) Representative images of immunofluorescence staining of Ki67 ( G , red) and CD31 ( I , red). ( H and J ) Quantitative analyses of immunofluorescence staining of Ki67 ( H ) and quantification of mean vessel density, branch points, and vessel length based on CD31 staining ( J ) in tumor sections of Panc1 xenograft (with or without PaSC co-implantation) mice treated with vehicle or ProAgio. Quantification of Ki67 staining is presented as the percentage of Ki67 + nuclei per view field (upper 3 panels, with PaSC co-implantation; bottom 3 panels, without PaSC co-implantation). ( C , G , and I ) Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI) (blue). ( A and B ) Error bars represent means ± SEM. ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001, ∗∗∗∗ P < .0001. n.s., denotes not significant.

Article Snippet: Sirius red, Masson trichrome, and Oil red O staining were performed using kits obtained from IHC WORLD (Woodstock, MD) by following the vendor’s instructions.

Techniques: Immunofluorescence, Staining

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Modulation of Cancer-Associated Fibrotic Stroma by An Integrin α v β 3 Targeting Protein for Pancreatic Cancer Treatment

doi: 10.1016/j.jcmgh.2020.08.004

Figure Lengend Snippet: ProAgio depletes CAPaSC by targeting integrin α V β 3 and resorbs tumor collagen. ( A ) Endogenous alleles of KrasG12D and Trp53R172H are conditionally activated in the pancreata of LSL-KrasG12D/+ ; LSL-Trp53R172H/+ ; Pdx-1-Cre (triple mutant) mice. ( B ) Specific polymerase chain reaction analysis of genomic DNA of generated GEM-KPC mice. ( C and D ) Kaplan–Meier survival analysis of GEM-KPC (median survival: vehicle, 20 days; ProAgio, 51 days) ( A ), and OrKPC ( B ) mice treated with vehicle or 10 mg/kg ProAgio (median survival: vehicle, 31 days; ProAgio, 39 days) (10 daily doses + 10 alternate day doses). Treatment was started after 13 weeks of age in GEM-KPC mice in panel C , or 30 days after tumor inoculation in panel D . The numbers in parentheses indicate the group size. ( E ) Representative images of Sirius red (red) and Masson trichrome (blue) staining and IHC staining of α-SMA of tumor sections from GEM-KPC mice treated with vehicle or 10 mg/kg ProAgio at different time points (R x 20, at end of 20 doses; End stage, end of survival experiments). ( F and G ) Quantitative analyses of Sirius red staining (presented as Sirius red + area %) ( F ) and αSMA + staining area % ( G ) in tumor sections of vehicle or ProAgio-treated (10 mg/kg) GEM-KPC mice. A higher staining of Sirius red was observed in vehicle-treated tumors on day 80 (R x 20 doses) than ProAgio-treated tumors on day 110 (end stage) in GEM-KPC mice ( P < .001; n = 7–9). ( H ) Representative images of Sirius red (Sirius red) and IHC staining of α-SMA in tumor sections of vehicle or ProAgio-treated (10 mg/kg) OrKPC mice at the end of 20 doses (R x 20 doses). ( I ) mRNA levels of α-SMA ( left panel ) and FAP ( right panel ) in the tumor tissue lysates of GEM-KPC mice treated with vehicle or ProAgio (n = 4) were analyzed by quantitative reverse-transcription polymerase chain reaction. The mRNA levels in the tumor extracts were normalized to mRNA level of β-actin. ( G ) Error bars represent means ± SEM. ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001, ∗∗∗∗ P < .0001. n.s., denotes not significant; WT, wild-type.

Article Snippet: Sirius red, Masson trichrome, and Oil red O staining were performed using kits obtained from IHC WORLD (Woodstock, MD) by following the vendor’s instructions.

Techniques: Mutagenesis, Polymerase Chain Reaction, Generated, Staining, Immunohistochemistry, Reverse Transcription

Journal: Signal Transduction and Targeted Therapy

Article Title: FGF1 ΔHBS prevents diabetic cardiomyopathy by maintaining mitochondrial homeostasis and reducing oxidative stress via AMPK/Nur77 suppression

doi: 10.1038/s41392-021-00542-2

Figure Lengend Snippet: FGF1 ΔHBS prevents cardiac dysfunction and remodeling in db/db mice. a – e db/db mice were treated with FGF1 ΔHBS (0.5 mg/kg body weight) or vehicle every other day for 16 weeks, and littermate db/m mice served as controls. a Echocardiographic parameters. n = 6. b Representative images of hematoxylin-eosin (H&E), WGA, Masson’s trichrome, Sirius red staining in cardiac tissues. c Quantification of myocyte area and cardiac fibrosis area in WGA, Masson’s trichrome and Sirius Red staining. n = 6. d Western blot analysis of collagen I (COL 1), collagen III (COL 3), myosin heavy chain (MyHC), cleaved caspase 3 (c-caspase 3) and transforming growth factor β1 (TGF-β1) in cardiac tissues; GAPDH was a loading control. e Densitometric quantification of western blots shown in d . n = 6. Data were mean ± SEM; * P < 0.05, ** P < 0.01, *** P < 0.001 in a , c ; *** P < 0.001 vs. db/m , ### P < 0.001 vs. db/db in e

Article Snippet: After dehydration, sections were stained with hematoxylin and eosin (H&E), Masson trichrome staining (Beyotime Biotech, Nantong, China) and Sirius red staining kits (Beyotime Biotech, Nantong, China) per manufacturer’s instruction.

Techniques: Staining, Western Blot