Supporting data for "SARS-CoV-2 Infection and COVID-19 mRNA Vaccination in Aged and Diet-Induced Obese Mouse Models"

1: Young, aged, lean and DIO (diet-induced obese) mice were intranasal inoculation with  Alpha or Omicron diluted in 20μl phosphate-buffered saline (PBS), body weight of the infected mice were monitored for a continued 14 days,.

2: RNA extracted from mice nasal turbinate (NT) and lung samples with Alpha or Omicron infection were extracted with a MiniBEST Universal RNA extraction kit (Takara Bio Inc. Shiga, Japan). RNA-dependent RNA polymerase (RdRp) gene copies of SARS-CoV-2 and cytokines were detected with a QuantiNova Probe RT-PCR Kit (208354, Qiagen), real-time RT PCR with corresponding primers quantified house-keeping gene β-actin for normalization were conducted by a LightCycler 96 system (Roche Applied Sciences, Indianapolis, USA) 

 3: Haematoxylin and Eosin (H&E) staining of sections (4μm) with formalin-fixed and paraffin-embedding in slides were observed for determine the changes of histopathology. For the expression of SARS-CoV-2 antigen, slides of NT and Lung tissues were stained with an in-house rabbit anti SARS-COV-2 nucleocapsid protein (NP) and then stained with a secondary antibody of FITC- conjugated goat anti rabbit IgG (Thermo Fisher Scientific, Waltham, MA, USA). Representative images were captured by using a microscope of Olympus BX53 semi-motorized bright-field or fluorescence with software of OLYMPUS Cell Sense.