Fig 1: Extracellular matrix protein deposition in BoNT-treated muscle-derived fibroblast. (A) Gene expression profiles of COL1A1, COL3A1, COL4A1, COL5A1, COL5A2, COL8A1 and COL11A1 expressed by fibroblasts extracted from the MG muscle in CP at t0 (immediately before in vivo BoNT administration, n = 10), CP at t1 (3 months after in vivo BoNT administration, n = 6), CP at t2 (6 months after in vivo BoNT administration, n = 9) and in TD children in the same age range (n = 7) exposed to 20 IU/mL BoNT for 48 h (for a visual representation, see also Figure S3A). Each data point represents individual values expressed as DCt and normalized for the housekeeping gene (bACT). Values are reported as mean ± SEM. Data were analyzed by two-way ANOVA with Tukey post hoc test. (B) WB and (C) protein level quantification for COL5A1, COL1A1 and COL3A1 normalized for aTUB expressed by fibroblasts extracted from TD and GMFCS-I CP samples untreated or treated with 20 IU/mL BoNT for 48 h. (D) WB and (E) protein level quantification for COL5A1, COL1A1 and COL3A1 normalized for aTUB expressed by fibroblasts extracted from TD and GMFCS-II CP samples untreated or treated with 20 IU/mL BoNT for 48 h. The same color highlights cells from the same sample. Different shades of green indicate GMFCS-I samples, of blue indicate GMFCS-II samples. Data were obtained from independent experiments, and values are reported as mean ± SD. Data were analyzed via one-way ANOVA with a Tukey post hoc test: * p < 0.05. CP vs. TD. (F) WB analysis and (G) protein level quantification for FN1, COL1A1 and COL3A1 normalized for aTUB expressed by untreated fibroblasts extracted from TD and GMFCS-I CP samples at t0 (immediately before in vivo BoNT administration), t1 (3 months after in vivo BoNT administration) or t2 (6 months after in vivo BoNT administration). (H) WB and (I) protein-level quantification for FN1, COL1A1 and COL3A1 normalized for aTUB expressed by untreated fibroblasts extracted from TD and GMFCS-II CP samples at t0 (immediately before in vivo BoNT administration), t1 (3 months after in vivo BoNT administration) or t2 (6 months after in vivo BoNT administration). Untreated samples are indicated by round symbols, while treated samples are represented by square symbols. The same color highlights cells from the same sample. Different shades of green indicate GMFCS-I samples, of blue represent GMFCS-II samples. Data were obtained from independent experiments, and values are reported as mean ± SD. Data were analyzed via one-way ANOVA with a Tukey post hoc test: * p < 0.05 for CP vs. TD; $$ p < 0.01 for CP t1 vs. CP t0.