Supplementary Table 1 from Resveratrol Induces Differentiation Markers Expression in Anaplastic Thyroid Carcinoma via Activation of Notch1 Signaling and Suppresses Cell Growth

PDF file - 44K, Primer sequences for quantitative real-time PCR.</p

Anti-HER2 mAb production in fed-batch cell culture.

(a) CHO/IgG (anti-HER2 mAb) cell growth in shaker flask (SF125) and 2-L stirred-tank bioreactor (BRX). Red rhombus: SF125 with working volume of 30 mL, Temp 37 oC, agitation 130 rpm, and CO2 5%. Blue square: BRX with working volume of 1 liter, Temp 37 oC, agitation 70 rpm, and DO 70%. (b) Anti-HER2 mAb production.</p

Summary of fed-batch cell culture in bioreactor.

Summary of fed-batch cell culture in bioreactor.</p

Figure S2 from Notch1 Signaling Regulates the Aggressiveness of Differentiated Thyroid Cancer and Inhibits SERPINE1 Expression

Scratch of a gap for wound healing assay was generated and closure of the gap was evaluated 24 hours later in FTC236-Notch1 cells with or without doxycycline treatment. The cells with NICD induction by doxycycline exhibited slower closure of the gap than those without NICD induction. Images were taken immediately after scratching the cultures 0 hr and 24hr later. Representative images at 0 hr and 24 hr are shown in the upper panel. The bar graph represents the average percentage of the gap closure. Each bar represents mean{plus minus} SEM; "Dox" indicates doxycycline; ** indicates p< 0.01; Scale bar:200µm.</p

Figure S4 from Notch1 Signaling Regulates the Aggressiveness of Differentiated Thyroid Cancer and Inhibits SERPINE1 Expression

SERPINE1 expression detected by immunohistochemistry in FTC236-Notch1 xenografts with or without Notch1 induction. (A) and (B). Two representative tumors from the group without Notch1 induction (Dox-); (C) and (D). Two representative tumors from the group with Notch1 induction (Dox+). Weaker staining of SERPINE1 (1+) is shown in the Dox+ tumors compared with 3+ staining in Dox- tumors. "Dox" indicates doxycycline; Scale bar: 50µm.</p

Summary of ADC cytotoxicity assay.

Summary of ADC cytotoxicity assay.</p

Evaluation of ADCs constructed in different production processes.

(a) Anti-cancer toxicity of free drugs. Blue triangle: MMAE, Red square: DM1, Green circle: mAb. (b) SDS-PAGE of ADCs. M: marker, 0: purified mAb, 1–4: lysine-based DM1-carrying ADCs (named as DM1-ADC1-4) with drug:linker:mAb ratio of 4:4:1, 8:4:1, 8:4:1, and 16:8:1, respectively. DM1-ADC1 and ADC2 were purified using G25 column, and DM1-ADC3 and ADC4 were purified using protein A column. 5–8: Cysteine-based ADCs. 5: ADC from sequential conjugation with rebridging linker. 6: ADC from sequential conjugation with traditional linker. 7: ADC from in situ conjugation with rebridging linker (named as MMAE-ADC1). 8: ADC from in situ conjugation with traditional linker (named as MMAE-ADC2). ADC samples were loaded to SDS-PAGE gel with 2 μg/well. (c) Anti-cancer toxicity of lysine-based anti-HER2 mAb-DM1 ADCs. Blue triangle: DM1-ADC1, Red square: DM1-ADC2, Green circle: DM1-ADC3, Purple rhombus: DM1-ADC4. (d) Anti-cancer toxicity of anti-HER2 mAb-MMAE ADCs. Blue triangle: MMAE-ADC1, Red square: MMAE-ADC2.</p

Supplementary Table S1 from Notch1 Signaling Regulates the Aggressiveness of Differentiated Thyroid Cancer and Inhibits SERPINE1 Expression

Sequences of each primer for real-time quantitative PCR</p

The diagram of ADC construction.

(a) Lysine-based conjugation of mAb-DM1: 1) mAb modification by cross linker Sulfo-SMCC, and 2) conjugation of DM1 with the purified mAb-SMCC. (b) Cysteine-based conjugation of mAb-MMAE using traditional linker: 1) synthesis of Mc-Val-Cit-PABC-PNP linker-MMAE payload, and 2) conjugation of payload with the DTT/TCEP reduced mAb. (c) Cysteine-based conjugation of mAb-MMAE using rebridging linker: 1) synthesis of rebridging linker, 2) synthesis of rebridging linker-MMAE payload, and 3) conjugation of payload with the DTT/TCEP reduced mAb. (d) Structures of free drugs MMAE and DM1.</p

Figure S1 from Notch1 Signaling Regulates the Aggressiveness of Differentiated Thyroid Cancer and Inhibits SERPINE1 Expression

The correlation between age and expression levels of Notch1 in primary DTC samples (A), contra-lateral non-tumor tissues (B) and tissue of benign thyroid disease (C).(A) Scatter plot of negative linear correlation between Notch1 levels and age in primary DTC samples (R = -0.495). (B)-(C) Significant linear correlation was not observed between age and Notch1 levels in the non-tumor tissues or benign thyroid lesions. Pearson's correlation test was used to compute the correlation significance between the 2 variables.</p