| Description |
To characterize the transcriptome landscape of Krt32 knockout and wildtype mice dermal response to TNF-α Stimulation, six C57BL/6J mice, each of Krt32 wild-type and knockout (KO), were subcutaneously injected with TNF-α at a dose of 6 ug/kg/day body weight into their shaved dorsal skin for a duration of 48 hours. After two days of TNF-α treatment, the dorsal skin of Krt32(-/-) mice exhibited pronounced thickening and extensive yellow scaling, resembling the cutaneous manifestations observed in human patients with pityriasis rubra pilaris (PRP). Furthermore, Krt32(-/-) mice exhibited a thickened epidermis, hyperkeratosis with focal parakeratosis, and mild lymphocyte infiltration in the superficial dermis compared to their wild-type counterparts. These observations were confirmed by hematoxylin-eosin staining of dorsal skin lesion sections. Next, the dorsal skin was separated using a scalpel (tissue size of 0.5cm x 0.5cm) and homogenized in RNA lysate reagent (LS1040, Promega) with the assistance of Gentle MACS Octo Dissociator (Miltenyi Biotec, Bergisch Gladbach, Germany). RNA quality was examined by gel electrophoresis and with Qubit (Thermo, Waltham, MA, USA). Sequencing was carried out using the Illumina Novaseq 6000 instrument by the commercial service of Genergy Biotechnology Co., Ltd. (Shanghai, China). |
