*5.12. Statistical Analysis*

Data are presented as means ± standard error (SEM) and analyzed by Graph Pad Prism, version 6.0 for Windows (Graph Pad Software, San Diego, CA, USA). One-way ANOVA test and multiple comparisons by Tukey HSD were used for comparisons of one variable in more than two groups. For comparisons of two or more variables, twoway ANOVA followed by Tukey HSD were used. For all tests, the values *p* < 0.05 were considered significant. The assays were conducted in duplicate and repeated at least twice in independent days.

**Supplementary Materials:** The following are available online at https://www.mdpi.com/article/ 10.3390/toxins13120906/s1, Figure S1. Differentiation of THP-1 macrophages. (A) Human premonocytes of the THP-1 lineage were cultured in 75 cm<sup>3</sup> bottles containing supplemented RPMI medium. The cells were kept in an incubator at 37 ◦C and 5% CO2, at a concentration of 2–8 <sup>×</sup> <sup>10</sup><sup>5</sup> cells/mL. (B) Macrophages were differentiated from THP-1 pre-monocytes with 100 ng/mL PMA for 3 days, followed by a 4-day rest period in the absence of PMA. 200× magnification. (C) CD11b expression in differentiated THP-1 macrophages. Adherent THP-1 macrophages differentiated with PMA and pre-monocytes in suspension were analyzed for CD11b expression by flow cytometry. Result expressed as mean of duplicates ± SEM and analyzed by two-way ANOVA followed by Tukey's post-test. (\*) Significant difference in relation to the respective controls (*p* < 0.0001). Figure S2. Cell cytotoxicity. Differentiated THP-1 macrophages was treated with 0.5–15 μg/well of BaV or with 0.5 and 1 μg/well Kn-Ba for 24 h, 48 h, and 72 h, and cell viability was evaluated by the quantification of lactate dehydrogenase (LDH) enzyme in cell-free supernatants. Results expressed as mean of duplicates ± SEM and analyzed by one-way ANOVA followed by Tukey's post-test. (\*) Significant difference in relation to the respective control. *p* < 0.01 (\*\*), and *p* < 0.0001 (\*\*\*\*).

**Author Contributions:** Conceptualization, Â.A.A.M. and W.D.-d.-S.; methodology, Â.A.A.M., F.C.V.P. and W.D.-d.-S.); validation, Â.A.A.M., F.C.V.P. and W.D.-d.-S.; formal analysis, Â.A.A.M. and W.D.-d.-S.; investigation, Â.A.A.M. and F.C.M.; resources, W.D.-d.-S. and F.C.V.P.; data curation, Â.A.A.M., F.C.M., F.C.V.P. and W.D.-d.-S.; writing—original draft preparation, Â.A.A.M., F.C.V.P. and W.D.-d.-S.; writing—review and editing, Â.A.A.M., F.R.G., K.S.G., F.C.V.P. and W.D.-d.-S.; supervision, W.D.-d.-S. and F.C.V.P.; project administration, W.D.-d.-S.; funding acquisition, W.D.-d.-S. and F.C.V.P. All authors have read and agreed to the published version of the manuscript.

**Funding:** This research was funded by Coordination for the Improvement of Higher-Level Education Personnel—CAPES (Â.A.A.M.: fellowship Ph.D. PROEX; W.D.S.: 23038.000814/2011–83); São Paulo Research Foundation (FAPESP 2013/07467-1 and Fapesp 2019/20832-7) and the National Council for Scientific and Technological Development (*PROÁFRICA*, CNPq: 490048/2005-6).

**Institutional Review Board Statement:** Not applicable.

**Informed Consent Statement:** Not applicable.

**Acknowledgments:** The authors thank Denise Tambourgi for the Immunochemistry Laboratory structure and to Martin Wesley for English reviewing in the article.

**Conflicts of Interest:** The authors declare no conflict of interest.

#### **References**

