Table 4. Summary of fixation methods used in Figure 7.

Detailed protocols for the different fixation methods used in Figure 7. Method 1 is the protocol used in this study.

Method	Pre-fixation	Pre-perme- abilization	Fixation	Quenching	Post-perme- abilization	Primary antibody	Secondery anitbody	References (use for nm-resolution analysis)
Method 1			3% PFA in PHEM (37°C)		0.5% NP40	37°C for 90 min	37°C for 90 min	Suzuki et al., 2011 ;Suzuki et al., 2014  ;Tauchman et al., 2015; this study
Method 2	3% PFA in PHEM (37°C)	0.5% triton in PHEM (37°C)	3% PFA in PHEM (37°C)		0.5% NP40	37°C for 90 min	37°C for 90 min	Wan et al. (2009); Varma et al., 2013
Method 3		1% triton in PHEM (37°C)	3% PFA in PHEM (37°C)		0.5% NP40	37°C for 90 min	37°C for 90 min
Method 4			1% GA in PHEM (37°C)	NaBH4	0.5% NP40	37°C for 90 min	37°C for 90 min
Method 5			1%GA w 1% triton in PHEM (37°C)	NaBH4	0.5% NP40	37°C for 90 min	37°C for 90 min	Magidson et al. (2016)
Method 6		1% triton in PHEM (37°C)	1% GA in PHEM (37°C)	NaBH4	0.5% NP40	37°C for 90 min	37°C for 90 min
Method 7		1% triton in PHEM (37°C)	1% GA w 3% PFA in PHEM (37°C)	NaBH4	0.5% NP40	37°C for 90 min	37°C for 90 min
Method 8			MeOH (−20°C)		0.5% NP40	37°C for 90 min	37°C for 90 min
Method 9			3%PFA w 1% triton in PHEM (37°C)		0.5% NP40	37°C for 90 min	37°C for 90 min	Magidson et al. (2016)