Anti cd147

The primary antibodies: anti-Cyclophilin A (1:1000 dilution, Abcam, ab126738), anti-CD147 (1:1000, ThermoFisher, MA1–19,201), anti-HSP70 (1:3000, Santa Cruz, sc-66,048), anti-TSG101 (1:1000 dilution, Abcam, ab125011), anti-HSP90B1 (1:1000 dilution, Cusabio, CSB-PA10887A0Rb), anti-GAPDH (1:500 dilution, EMD Millipore, MAB374), anti-Rab7 (1:1000 dilution, Cell signaling Technology, 9367), anti-Rab11 (1:250 dilution, ThermoFisher, 71–5300).
The secondary antibodies: anti-Rabbit IgG-DyLight 800 (1:10,000 dilution, ThermoFisher, SA5–35,571), anti-Mouse IgG-DyLight 680 (1:10,000 dilution, ThermoFisher, 35,519)
Reagents: Trypsin (Pierce Trypsin Protease, MS Grade, ThermoFisher, 90,057), Propidium Iodide (ThermoFisher, P1304MP), 5-(and −6)-Carboxyfluorescein Diacetate, Succinimidyl Ester, mixed isomers (5(6)-CFDA, SE) (ThermoFisher, C1157)

Induced Pluripotent Stem Cells-MNs were seeded on coverslips in a 24-well plate, cultured until differentiation, and infected as specified above. At 48 hpi, cells were fixed in PBS containing 4% PFA at RT for 10 min, followed by permeabilization with 0,1% TritonX-100 in PBS for 10 min. Cells were treated with 1% BSA in PBS for blocking at RT for 1 h, and incubated at 4°C overnight with specific primary antibodies. The following primary antibodies were used: anti-beta III Tubulin (1:500, Abcam, Cambridge, UK), anti-SMI-312 (1:1000, Covance, Princetown, NJ, USA) and anti-ChAT (1:200, Chemicon) to assess iPSC-MN differentiation; anti-ACE2 (1:200, Prodotti Gianni), anti-CD147 (1:100, Thermo Fisher Scientific) anti-NRP1 (1:100, Thermo Fisher Scientific) and anti-N Nucleocapsid SARS-CoV-2 (1:1000, BEI Resources) to assess SARS-CoV-2 receptors and infection. Coverslips were then stained with secondary antibodies (Alexa Fluor 488 or 647, 1:500, Abcam) for 45 min at RT and mounted using a medium containing DAPI (Enzo Life Sciences, Milan, Italy). Confocal images were acquired on a TCS SP8 System equipped with a DMi8 inverted microscope and a HC PL APO 40 × /1.30 Oil CS2 (Leica Microsystems, Wetzlar, Germany) at a resolution of 1024 × 1024 pixels (single stack).