Originpro 2020b

For the fractional factorial design, statistical significance of the main effects was evaluated using analysis of variance (ANOVA) with a 95% confidence interval. In comparing the extraction efficiencies of pure water and aqueous methanol (MeOH:H₂O, 50% v/v), a t-test was used to assess whether the results of the two extractants were significantly different. Statistica (v. 13.5.0.17, TIBCO, Palo Alto, CA, USA) was used in generating the experimental design and processing the corresponding analytical results. Microsoft Excel (Microsoft, Redmond, WA, USA) was used in statistical treatment of data and calculation of other analytical figures of merit. OriginPro 2020b (v. 9.7.5.184, OriginLab, Northampton, MA, USA) was used in creating figures.

The spectra were analyzed using OriginPro 2020b (OriginLab Corporation, USA). The analysis included spectra baseline correction, smoothing using the Savitzky–Golay filter (polynomial order 2, widow size 35, SG 35) [82 (link)], normalization of spectra relative to Amide I band (ATR-FTIR), or deformation vibrations of CH₂ group, at 1450 cm⁻¹ (FT-Raman).
PCA was performed on the second derivative of the Amide I region of the spectra (1725–1590 cm⁻¹) using Scikit-learn Python package [83 ]. Matplotlib [84 (link)] Python package was used for visualization.

Electron microscopy was carried out as described previously (37 (link)). The purified carboxysomes (∼4 mg mL⁻¹) were stained with 3% uranyl acetate on carbon grids and then inspected with an FEI 120 kV Tecnai G2 Spirit BioTWIN transmission electron microscope (TEM) equipped with a Gatan Rio 16 camera. The diameters of carboxysomes were measured with ImageJ as described previously (37 (link)) and were statistically analyzed using OriginPro 2020b (OriginLab, MA).

In order to determine the hydrodynamic diameter of the sample, a Vasco Particle Size Analyzer from Cordouan Technologies (Pessac, France) was used. The parameters of the determination were laser power 100%, temperature 25 °C, number of channels 350, time interval 14 µs, DTC position down, and acquisition mode continuous. The measurements were performed in triplicate. The obtained data were plotted and analyzed with the Gauss non-linear curve fit using the OriginPro 2020b software (OriginLab Corporation, Northampton, MA, USA).
For the zeta potential measurement, a Wallis Zeta-potential Analyzer from Cordouan Technologies (Pesac, France) was used. The parameters of the measurement were medium resolution, temperature 25 °C, plastic cuvette, laser power 65%, electrode distance 5 mm, and Henry function Huckel. Ten measurements were performed for each sample.
For the DLS determination and the zeta potential measurements, a suspension was made by sonication (ultrasonic bath) using the synthesized magnetic powder/nanobioconjugates and distilled water.

PrOF NMR was performed as reported previously^{36 (link)}. Briefly, 150 µM 5FW LecA was recorded alone and in the presence of DMSO or 2–4 mM compounds in TBS/Ca²⁺ buffer. The difference in chemical shift perturbations of W42 in 5FW LecA free vs bound forms was followed to determine K_d values of compounds. The K_d values were calculated according to the one-site-binding model in Origin(Pro) 2020b (OriginLab Corp., USA) from two or three independent titrations. Ligand efficiency values were calculated according to Eq. (1) for a temperature of 310 K.

Chemometric analyses such as hierarchical cluster analysis (HCA), principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS–DA) were performed to systematically analyze the influence of tree age on the volatile component contents in ancient Platycladus orientalis leaves. Heatmap analysis and principal component analysis were generated using the Origin software for statistical and computing (Origin Pro 2020b, Origin Lab, USA). Orthogonal partial least squares discriminant analysis was performed using R (http://www.r-project.org/ (accessed on 15 November 2022)). Variance analysis was performed using SPSS 24.0 for Windows (SPSS Inc., Chicago, IL, USA).

To show statistical differences between samples, hierarchical clustering analysis (HCA) was performed with the first five principal components as input data calculated on the base of all measured parameters to minimize noise in the data. The first five components accounted for a sum of variances higher or equal to 98.75%. The cluster method was based on the group average. In addition, two-way ANOVA was performed to statistically evaluate the differences between different storage conditions and the shape of hops. Data were analyzed using the OriginPro^® 2020b (OriginLab Corporation, Northampton, MA, USA) software package.