Eif3h

Murine palatal shelf tissues of E13.5, E14.5, and E16.5 were dewaxed in xylene, and then dehydrated in ethanol. The tissues were then incubated in solutions containing specific primary antibodies: EIF3H (1:100, Santa Cruz, sc-271283), SRSF1 (1:100, Santa Cruz, sc-33652), and RBBP6 (1:100, Santa Cruz, sc-9962) (Table S3) followed by incubation in Alexa fluor 488-labeled goat anti-mouse secondary antibodies (1: 50, Beyotime, A0428).

Hematoxylin and eosin (H&E) staining was performed according to the manufacturer’s instructions. In brief, human sample tissues of NSCL/P were immersed in 4% paraformaldehyde for 48 h. Next, the fixed tissues were dehydrated, cleared, and embedded in paraffin wax. The paraffin blocks were cut into 4-μm-thick sections and stained with hematoxylin and eosin (H&E). Primary antibodies against the following proteins were used: EIF3H (1:100, Santa Cruz, sc-271283), SRSF1 (1:100, Santa Cruz, sc-33652), and RBBP6 (1:100, Santa Cruz, sc-9962) (Table S3). Briefly, tissues were fixed, dehydrated, embedded, and sectioned for each sample for all stains. Sections were incubated with primary antibodies, washed, and then incubated with appropriate secondary antibodies (MaxVision, kit-5020, China). Optical microscopy (Thermo Scientific, Wilmington, USA) was used to image stained sections. Semi-quantitative analysis was performed using Fiji v2.9.0 (NIH, Bethesda).