Cytochalasin d ab143484

For the flow cytometry analysis, anti-CD4 (GK1.5), anti-CD8α (53–6.7), anti-CD44 (IM7), anti-CD127 (A7R34), anti-CD69 (FN50), anti-Bcl-2 (BCL/10C4) were purchased from Biolegend. Anti-CD62L (MEL-14), anti-CD69 (H1.2F3), anti-CD5 (53–7.3), anti-CD45.1 (A20), anti-CD45.2 (104), mouse lgG1 κ isotype control (556027), anti-CD25 (PC61), Rat lgG2a isotype control (eBRG1) and purified anti-human-CD3 (OKT3) were purchased from BD Bioscience. Anti-Foxp3 (FJK-16s), anti-TCRβ (H57–597) and anti-CD3 (UCHT1) were purchased from eBioscience. Fluo-4,AM (F14217) was purchased from Invitrogen. Streptavidin (NEBN7021S) was purchased from Sigma.
For western analysis, anti-α-tubulin (sc-53029) and anti-Arp3 (sc15390) antibodies were purchased from Santa Cruz Biotechnology. Anti-Arpc2 (ab96779) was purchased from Abcam.
For immunofluorescence analysis, anti-Mouse TCR β-chain-488 (H57-597) and anti-Mouse CD3-488 (17A2) were purchased from Biolegend. Anti-CD3ζ (6B10.2) was purchased from Santa Cruz. Anti-EEA1 (C45B10), anti-Rab5 (C8B1) and anti-Rab11 (D4F5) were purchased from Cell Signaling Technology. Anti-LAMP1 (ab24170) was purchased from Abcam.
Cytochalasin D (ab143484) was purchased from Abcam. Staphylococcal enterotoxin E (ET404) was purchased from Toxin Technology. CELLTRACKER(TM) BLUE CMAC (C2110) was purchased from Invitrogen.

To prepare the Ca²⁺-free medium, Hank’s balanced salt solution (HBSS; 14175-095, Gibco) was supplemented with 20 mM hydroxyethylpiperazine ethane sulfonic acid (HEPES; Dojindo), 0.5 mM ethylene glycol-bis(2-aminoethylether)-N,N,N’,N’-tetraacetic acid (EGTA; E3889, Sigma), 1 mM magnesium chloride (MgCl₂; M8266, Sigma), and 1 mM magnesium sulfate (MgSO₄; 746452, Sigma) (Kim et al., 2015 (link)). To prepare the Ca²⁺ medium, HBSS was supplemented with 20 mM HEPES, 1 mM MgSO₄, and 1 mM calcium chloride (CaCl₂; 18230S0301, JUNSEI). The Ca²⁺ medium did not include EGTA, a calcium chelator, or MgCl₂, a supplemented ion. To represent normal conditions, CO₂-independent culture medium (18045088, Gibco) containing 0.5% (v/v) FBS, 4 mM l-glutamine (LS002-1, Welgene), 100 U/ml penicillin, and 100 μg/ml streptomycin was used. Yoda1 (SML1558), methyl-β-cyclodextrin (MβCD; 332615), ML-7 (I2764), gadolinium (Ⅲ) chloride hexahydrate (G7535), and thapsigargin (T9033) were purchased from Sigma-Aldrich. Nocodazole (S2775) was purchased from Selleckchem. Cytochalasin D (ab143484) was obtained from Abcam.