Host Cell Copper Transporters CTR1 and ATP7A are important for Influenza A virus replication

Table 1 Elemental analysis of total intracellular copper in A549 cells by ICP-MS

A549 Treatment^a	Protein BCA^b ug/ul	Ionized Cu²⁺ ICP-MS^c ug/g	Intracellular Cu²⁺ Ratio to Control^d
Media	0.663	4.0	1.0
10 μM TTM	0.686	1.2	0.3
50 μM CuCl2	0.144	12.9	14.8
Negative Control siRNA	0.680	2.7	1.0
CTR1 esiRNA	0.472	1.6	0.9
ATP7A esiRNA	0.502	18.2	9.1

^aA549 cells were untreated (Media), treated with 50 μM CuCl₂ or 10 μM ammonium tetrathiomolybdate (TTM), or transfected with indicated siRNA for 24 hours, washed in PBS and lysed in RIPA buffer
^bProtein measured by BCA assay
^cElemental analysis of total intracellular copper (ionized to Cu²⁺) by inductively coupled plasma mass spectrometry (ICP-MS)
^dIntracellular Cu was normalized to protein content for control conditions of Media alone (χ² < 0.001), or Negative Control siRNA (χ² < 0.001), expressed as a ratio to each control condition, respectively; χ², chi-squared test against null model

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