Go 6983

Recombinant human epidermal growth factor (EGF) and C-X-C motif chemokine 12 (CXCL12) was acquired from Sigma-Aldrich (E5036 and SRP4391). Cytochalasin D (Cyto D) was obtained from MilliporeSigma (Burlington, MA, USA). Blebbistatin (Bleb) was from Aladdin (Shanghai, China). PMA (PKC activator; HY-18739), Go 6983 (PKC inhibitor; HY-13689) and LY-294002 (PI3K inhibitor; HY-10108) were obtained from MedChemExpress. The following antibodies were used: Rabbit anti-srGAP2 (Proteintech, #22519-1-AP); Rabbit anti-β-actin (CST, #4970); Mouse anti-actin (Abcam, #ab179467); Mouse anti-p-Ser (Santa, #sc-81514); Rabbit anti-PKCα (Abcam, #ab32376); Rabbit anti-syndecan-4 (Abcam, #ab74139); Mouse anti-yndecan-4 (Santa, #sc-12766); Rabbit anti-FMNL1 (Proteintech, #27834-1-AP); Rabbit anti-ATP1A1 (Proteintech, #14418-1-AP); Rabbit anti-FMNL3 (Sigma, #PA5-46742) and Mouse anti-Talin 1 (Abcam, #ab108480).

Huh1, Huh6, SNU878, JHH-7 and Huh7 cells were obtained from the Institute of Biochemistry and Cell Biology, Chinese Academy of Science, China. HepG2, Hep3B, PLC/PRF/5, SNU387, SNU423, SNU398, and SNU449 cells were purchased from ATCC (Manassas, VA, USA). MHCC97H, HCCLM3, and HCCLM6 cells were kindly provided by Dr. Tang ZY (Liver Cancer Institute, Zhongshan Hospital, Fudan University, Shanghai, China). Cells were cultured in DMEM or 1640 medium with 10% FBS, 100 μg/ml penicillin, and 100 μg/ml streptomycin at 37 °C in a 5% CO2 incubator. All the cell lines were certified by the STR.
The ERK inhibitor SCH772984 (HY-50846), JNK inhibitor SP600125 (HY-12041), P38 inhibitor SB203580 (HY-10256), PI3K inhibitor LY294002 (HY-10108), PKC inhibitor GO6983 (HY-13689), recombinant human basic FGF (HY-P7004), FGFR1 inhibitor PD173074 (HY10321) and ACLY inhibitor ETC-1002 (HY-12357) were purchased from MedChemExpress.

Remdesivir (catalog no. SF-1193) was purchased from Beyotime (Shanghai, China). Go 6983 (catalog no. HY-13689), bisindolylmaleimide I (catalog no. HY-13867), enzastaurin (catalog no. HY-10342), sotrastaurin (catalog no. HY-10343), rottlerin (catalog no. HY-18980), and ebselen (catalog no. HY-13867) were purchased from MedChemExpress (MCE) (Shanghai, China).

Remdesivir (catalog no. SF-1193) was purchased from Beyotime (Shanghai, China). Go 6983 (catalog no. HY-13689), bisindolylmaleimide I (catalog no. HY-13867), enzastaurin (catalog no. HY-10342), sotrastaurin (catalog no. HY-10343), rottlerin (catalog no. HY-18980), and ebselen (catalog no. HY-13867) were purchased from MedChemExpress (MCE) (Shanghai, China).

Primary NSCs cultures were prepared from SVZ of newborn Seipin KO and WT mice euthanized by cervical cord dislocation following the protocol established previously [15 (link)]. Briefly, cells were plated at 1 × 10⁵ cells/mL and cultured in DMEM-F12 proliferation medium supplemented with 2% B27 (Invitrogen, Stockholm, Sweden), 20 ng/mL epidermal growth factor (EGF; PeroTech, Rocky Hill, NJ, USA), and 20 ng/mL basic fibroblast growth factor (bFGF; PeroTech) in a humidified incubator at 37 °C with 5% CO₂. After a week of culture, neurospheres were gained and digested with Accutase Cell Detachment Solution (Gibco) to obtain single cell suspension. Then NSCs were plated onto the coverslips pre-coated with 50 μg/ mL poly-L-ornithine and 20 μg/mL laminin (Sigma-Aldrich, Saint Louis, MO, USA) overnight. To measure the effect of DG on NSCs proliferation, cells were treated with 50 μM and 100 µM DiC8 (Aladdin Chemistry Co, Shanghai, China) or PKC inhibitor (Go6983; 140 nM; MedChemExpress, Monmouth Junction, NJ, USA) and cultured in DMEM supplemented with BrdU (10 μM) for 12 h for immunofluorescence analysis. For NSCs differentiation, the culture medium was replaced with differentiation medium containing 1% fetal bovine serum, 1% B27 supplement, DiC8, and/or Go6983. After three days, cells were harvested for immunostaining.