. 2023 Nov 8;12:e80749. doi: 10.7554/eLife.80749

Table 1. Side-by-side comparison of parameters and cell subtype identification criteria in the current study and Jiang et al., 2018.

Citations provided in ‘Materials and methods’.

	Current study		Jiang et al., 2018
	M2	M4	M2	M4
Mouse models	Opn4-GFP Trpc3^-/-; Trpc6^-/-; Trpc7^-/-	WT Trpc3^-/-; Trpc6^-/-; Trpc7^-/-	Opn4-tdTomato Primarily: Trpc6^-/-; Trpc7^-/- Figure 2: Trpc1^-/-; Trpc3^-/-; Trpc4^-/-; Trpc5^-/-; Trpc6^-/-; Trpc7^-/-	Opn4-tdTomato Primarily: Trpc6^-/-; Trpc7^-/- Figure 2: Trpc1^-/-; Trpc3^-/-; Trpc4^-/^-; Trpc5^-/-; Trpc6^-/-; Trpc7^-/-
Cell targeting ex vivo	Epifluorescence intensity	IR-DIC/soma size and ON-sustained light response	Epifluorescence intensity/soma size	Epifluorescence intensity/soma size
Subtype identity	During recording: Stratification analysis using Alexa 594 Neurobiotin fill for post-hoc morphological analysis Post recording: SMI-32 negative ON-stratification Large arbors with moderate branching Defining feature: M2 ipRGCs are the only ON stratifying ipRGC labeled in Opn4-GFP mice and they are SMI-32 negative.	During recording: Stratification analysis using Alexa 594 Neurobiotin fill for post hoc morphological analysis Post recording: SMI-32 positive ON stratification Highly branched arbors Defining feature: M4 ipRGCs are the only SMI-32 positive ipRGC, and are ON stratifying	Intracellular dye filling: Alexa 568 for morphological analysis of dendritic arbors (criteria unspecified)	Intracellular dye filling: Alexa 568 for morphological analysis of dendritic arbors (criteria unspecified)
Mice dark-adapted	Vernight		3 hr
Technique	Whole-cell voltage-clamp recording		Whole-cell voltage-clamp recording
Holding potential	–66 mV for all experiments unless otherwise mentioned		–66 mV for all experiments except Figure 4
Internal Solution (mM)	120 K-gluconate, 5 NaCl, 4 KCl, 10 HEPES, 2 EGTA, 4 ATP-Mg, 0.3 GTP-Na₂ and 7-Phosphocreatine-Tris, with the pH adjusted to 7.3 with KOH		120 K-gluconate, 5 NaCl, 4 KCl, 10 HEPES, 2 EGTA, 4 ATP-Mg, 0.3 GTP-Na₂ and 7-Phosphocreatine-Tris, with the pH adjusted to 7.3 with KOH
Synaptic Blockers	100 μM DNQX, 20 μM L-AP4, 100 μM picrotoxin, and 20 μM strychnine		20 μM DNQX, 50 μM AP5, 100 μM Hexamethonium, 100 μM picrotoxin, and 1 μM Strychnine
Recording Temperature	30–32°C		30–32°C
ZD7288 Conditions	Concentration: 50 μM Incubation time for effective HCN blockade: 5–8 min Incubation time driving off-target effects: 20 min		Concentration: 50 μM Incubation time: not reported
Light step	50 ms		200 ms
Light intensity	6.08 × 10¹⁵ photons · cm^–2 · s^–1 blue LED light (480 nm)		White light of an intensity equivalent to 1.75 × 10¹⁸ photons cm^–2 s^–1 of 480 nm light for melanopsin (conversion done by response-matching in the linear range)

IR-DIC, infrared differential interference contrast; ipRGCs, intrinsically photosensitive retinal ganglion cells.