Anti guinea pig alexa 647

Purified gametocytes were activated for 1–2 min and then activation was stopped by adding 4% formaldehyde. Sample preparation of P. berghei parasites for UExM was performed as previously described^{68 (link),69 (link)}, except that 4% formaldehyde (FA) was used as fixative^{43 (link)}. Fixed samples were then attached on a 12 mm round poly-d-lysine (A3890401, Gibco) coated coverslip for 10 min. Immuno-labelling was performed using primary antibodies against α-tubulin and β-tubulin (1:200 dilution, AA344 and AA345 from the Geneva antibody facility), anti-γ-tubulin antibody (1:500 dilution, Sigma T5192) and anti-HA antibody (3F10) (1:250 dilution, Roche). Secondary antibodies anti-guinea pig Alexa 647, anti-rabbit Alexa 405 and anti-rat Alexa 488 were used at dilutions 1:400 (Invitrogen). Atto 594 NHS-ester was used for bulk proteome labelling (Merck 08741). Images were acquired on a Leica TCS SP8 microscope, image analysis was performed using Fiji-Image J and Leica Application Suite X (LAS X) software.

Adipose tissues were harvested from perfused (4% paraformaldehyde) mice. Paraffin embedding and tissue sectioning was performed by the Molecular Pathology Core Facility at UTSW. Indirect immunofluorescence was performed as previously described (Vishvanath et al., 2016 (link)). Antibodies used for immunofluorescence include: anti-GFP 1:700 (Abcam ab13970), anti-perilipin 1:1500 (Fitzgerald 20R-PP004), anti-chicken Alexa 488 1:200 (Invitrogen), and anti-guinea pig Alexa 647 1:200 (Invitrogen). Hematoxylin and eosin staining was performed according to manufacturer’s instructions.