Table 2 Reagents

Danieau’s solution

17.4 mM NaCl, 0.21 mM KCl, 0.12 mM MgSO₄•7H₂O, 0.18 mM Ca(NO₃)₂, 1.5 mM Hepes

Fixation solution

4% PFA in PBS, pH 7

PBS

137 mM NaCl, 2.7 mM KCL, 10 mM Na₂HPO₄, 1.8 mM KH₂PO₄, pH 7.4

PBT 0.1%/0.01%

0.1%/0.01% Tween-20 in PBS

SSCT 0.01%

0.01% Tween-20, 15 mM NaCl, 1.5 mM TriNaCitratdihydrate, pH 7

Negative control dapB (cat.# 310043)

Channel 1, ready-to-use solution

Advanced Cell Diagnostics (ACD), Hayward, CA, USA

Target probe cxcl12a (cat.# 406481)

Channel 1: ready-to-use solution

ACD, Hayward, CA, USA

Target probe myoD (cat.# 402461-C2)

Channel 2: 50× solution

ACD, Hayward, CA, USA

Target probe vasa (cat.# 407271-C3)

Channel 3: 50× solution

ACD, Hayward, CA, USA

Target probe cxcr4b (cat.# 418121-C3)

Channel 3: 50× solution

ACD, Hayward, CA, USA

Target probe egfp (cat.# 400281)

Channel 1: ready-to-use solution

ACD, Hayward, CA, USA

Target probe nanos (cat.# 404521-C2)

Channel 2: 50× solution

ACD, Hayward, CA, USA

Pretreat 3 in RNAscope pretreatment kit (cat.# 320842)

Pretreat 3 ready-to-use solution

ACD, Hayward, CA, USA

RNAscope fluorescent multiplex detection kit (cat.# 320851)

Amp1, Amp2, Amp3, Amp4 AltA, Amp4 AltB, Amp4 AltC, Pretreat 3, DAPI ready-to-use solutions, Wash buffer provided in the kit is not used and is replaced with 0.2× SSCT (0.01% Tween-20)

ACD, Hayward, CA, USA

1% or 4% LMP

1% or 4% low melting point agarose in Danieau’s solution

Invitrogen, Paisley, UK

Hoechst 33342 (cat.# H1399)

Use at a 1:10000 dilution in 0.2× SSCT (0.01% Tween-20)

Invitrogen, Paisley, UK