arithmetic mean intensity  (Carl Zeiss)

Journal: Scientific Reports

Article Title: Regulation of serine palmitoyl-transferase and Rac1–Nox2 signaling in diabetic retinopathy

doi: 10.1038/s41598-022-20243-2

Figure Lengend Snippet: Inhibition of SPT and Rac1–Nox2–ROS signaling in HRECs. ( a ) SPT mRNA was quantified by qRT-PCR using β-actin as a house keeping gene. ( b ) Nox2 by using lucigenin as an electron acceptor and NADPH as a substrate and ( c ) Rac1 activity was determined by G-LISA colorimetric assay kit. ( d ) ROS levels were quantified by a fluorescence method using DCHFDA. ( e ) Transfection efficiency of SPT -siRNA was determined by quantifying the relative mRNA of SPT . Each measurement was made in duplicate, and the values are represented as mean ± SD from 3–4 cell preparations. NG and HG = 5 mM or 20 mM d -glucose; HG + PA = 20 mM d -glucose + palmitic acid; HG/ SPT -si and HG + PA/ SPT -si = SPT -siRNA transfected cells in 20 mM d -glucose or 20 mM d -glucose + palmitic acid; HG + PA/Myr = 20 mM d -glucose + palmitic acid + Myriocin; HG/SC = SPT -scrambled control RNA transfected cells in 20 mM d -glucose; L-Gl = 20 mM l -glucose. *p < 0.05 vs NG, # p < 0.05 vs HG and # **p < 0.05 vs HG + PA.

Article Snippet: The ‘Arithmetic mean intensity’ (AMI) of Rac1 and Nox2, and Pearson correlation coefficient between them were quantified using Zeiss software module .

Techniques: Inhibition, Quantitative RT-PCR, Activity Assay, Colorimetric Assay, Fluorescence, Transfection, Control

Journal: Scientific Reports

Article Title: Regulation of serine palmitoyl-transferase and Rac1–Nox2 signaling in diabetic retinopathy

doi: 10.1038/s41598-022-20243-2

Figure Lengend Snippet: Rac1–Nox2 colocalization and inhibition of SPT. ( a ) Rac1 and Nox2 were determined by immunofluorescence using DyLight green (green)-conjugated and Texas Red (red)-conjugated secondary antibodies, respectively. The line marker represents 20µm. ‘Arithmetic mean intensity’ (AMI) of ( b ) Nox2 and ( c ) Rac1 was quantified by Zeiss software, from 5 to 8 images/group. ( d ) Zeiss software module was used to calculate Pearson’s correlation coefficient between Rac1 and Nox2. The values in the graph are mean ± SD, obtained from 3–4 different cell preparations with multiple cells analyzed in each preparation. *p < 0.05 vs NG, # p < 0.05 vs HG and # **p < 0.05 vs HG + PA.

Article Snippet: The ‘Arithmetic mean intensity’ (AMI) of Rac1 and Nox2, and Pearson correlation coefficient between them were quantified using Zeiss software module .

Techniques: Inhibition, Immunofluorescence, Marker, Software

Journal: Scientific Reports

Article Title: Regulation of serine palmitoyl-transferase and Rac1–Nox2 signaling in diabetic retinopathy

doi: 10.1038/s41598-022-20243-2

Figure Lengend Snippet: Rac1–Nox2–ROS signaling and SPT inhibition in retinal microvessels from diabetic mice. Retinal microvessels were analyzed for ( a ) Rac1 activity, ( b ) Nox2 activity, and ( c ) ROS by DCHFDA method. Each measurement was made in duplicate, and the values are means ± SD of 5–7 mice/group. Norm = normal mice on regular chow; Diab = diabetic mice on regular chow; HF-SD = diabetic mice on high fat chow; Diab /SPT -si or HF-SD/ SPT -si = SPT -siRNA administered diabetic mice on regular chow or high fat chow, respectively. *p < 0.05 vs Norm, # p < 0.05 vs Diab and # **p < 0.05 vs HF-SD.

Article Snippet: The ‘Arithmetic mean intensity’ (AMI) of Rac1 and Nox2, and Pearson correlation coefficient between them were quantified using Zeiss software module .

Techniques: Inhibition, Activity Assay